NOTEBOOK
December
9.12.-13.12. We filled a survey to become next generation of ATOMS members.
Waiting in excitement and wondering who will be the members of this year’s team.
22.12. Aaaaand the results of the interviews we made are announced! Now we’re a crowded team of 25 people. Let’s see what’ll happen next.
26.12. Had our first meeting and our first topic was: sponsorship.
January
We had training classes at 08.01-10.01. January. Now it is time to learn what iGEM wants from us.
10.01.-23.01. We made researches about previous iGEM teams, wondering who did what.
23.01.-28.01. Hooray! It’s holiday! Home sweet home.
It’s time to find our own project. After now, we started to have meetings every Monday and Thursday, becase we need to work a lot for finding a new project. Also, still trying to find sponsors.
February
01.02. As we are ATOMS, those Sundays needed to be filled too! We made subgroups in our team and started to search previous Grand Prized-Teams. Now every group will present a team at the meetings.
We have just built ATOMS gene library.
02.02. We decided to attend some competitions due to the lack of sponsors. We attended ‘illnesses with imagery’. Talented people is a must in a team.
13.02. Drawings were done and sent. We believe we will be successful. We are still doing researches in full swing.
March
05.03. Today they filmed our school, and our laboratory so. We had so much fun while they were shooting us!
11.03. The art competiton’s results are just announced. Disappointment. There was not even a winner of first place. We convinced ourselves with thinking they don’t have a sense of art.
14.03. We made a biobrcik design workshop till we see the sunlight! At the late times of night, every group explained their biobrick designs. Now everyone knows what a promoter is :)
17.03. Lab-cleaning party! We don’t wanna be contaminated.
April
05.04. We bid farewell to our teammate Furkan Beştepe. He went to USA.
10.04. Happy birthday Gülnihal! By the way your birthday cake was yummy.
12.04. Daytime wasn’t enough so we started to have meetings in the nights! After working a lot, we deserved eating a delicious meal. Today we also started to search about Toehold.
13.04. We are still learnig about Toehold. It has a potential to be used.
27.04. Fun at the lab.
30.04. ATOMS Ladieas worked on a universal blood idea till the morning.
May
03.05. We must hurry up to find a project. Camp time at Asya Thermal Hotel.
04.05. Why there isn’t anyone in the lab?
06.05. Finally found our project idea! ULCER AND CANCER
09.05. Perfecting our cancer switch system.
15.05. Presented our project to the dean Mehmet Gündüz and the whole genetic department of our faculty! He treated us with Turkish tea and snacks.
24.05. We held a meet-up with other Turkish iGEM teams! Time to make some presentations.
June
This month we designed our genes and studied for our exams at the same time. In short it was a tough week.
Week 1Gradient PCR from pCAGGS | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR fwd | VR rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 0.5 ul | 0.5 ul | 0.5 ul | 0.2 ul | 16.3 ul | 2.0 ul | 25.0 ul |
9X | 22.5 ul | 22.5 ul | 4.5 ul | 4.5 ul | 4.5 ul | 1.8 ul | 146.7 ul | 18.0 ul | 225.0 ul |
57-64˚C
Results weren’t matching with expected results, experiment will be repeated.
(30.06.2015)
Gradient PCR from pCAGGS | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR fwd | VR rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 0.5 ul | 0.5 ul | 0.5 ul | 0.2 ul | 16.3 ul | 2.0 ul | 25.0 ul |
9X | 22.5 ul | 22.5 ul | 4.5 ul | 4.5 ul | 4.5 ul | 1.8 ul | 146.7 ul | 18.0 ul | 225.0 ul |
52-59˚C
Results weren’t matching with expected results, experiment will be repeated.
August
11.08. We started designing our wiki.
18.08.2015 We got a little tired of doing experiments, so treated ourselves with an awesome team dinner at a fish restaurant.
22.08.2015 A team dinner again, but this time our dear professors Mehmet and Esra Gündüz hosted us at their house.
29.08.2015 Wiki’s tour part is drawn, now it’s time to color these drawings!
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/1/2015 6:03:37 PM | -0.1 | ng/ul | -0.003 | -0.009 | 0.30 | 0.12 | DNA | 50.00 |
2 | psb1c3-A | biospec | 8/1/2015 6:06:00 PM | 207.0 | ng/ul | 4.140 | 2.184 | 1.90 | 2.11 | DNA | 50.00 |
3 | psb1c3-B | biospec | 8/1/2015 6:07:07 PM | 209.0 | ng/ul | 4.179 | 2.210 | 1.89 | 2.19 | DNA | 50.00 |
4 | psb1c3-C | biospec | 8/1/2015 6:08:05 PM | 157.3 | ng/ul | 3.147 | 1.664 | 1.89 | 2.09 | DNA | 50.00 |
5 | psb1c3-D | biospec | 8/1/2015 6:08:58 PM | 242.5 | ng/ul | 4.851 | 2.548 | 1.90 | 2.11 | DNA | 50.00 |
6 | psb1c3-E | biospec | 8/1/2015 6:09:42 PM | 116.4 | ng/ul | 2.329 | 1.215 | 1.92 | 2.17 | DNA | 50.00 |
7 | psb1c3-F | biospec | 8/1/2015 6:10:28 PM | 236.4 | ng/ul | 4.729 | 2.503 | 1.89 | 2.17 | DNA | 50.00 |
8 | psb1c3-71 | biospec | 8/1/2015 6:11:33 PM | 111.7 | ng/ul | 2.233 | 1.175 | 1.90 | 2.10 | DNA | 50.00 |
9 | psb1c3-72 | biospec | 8/1/2015 6:12:14 PM | 106.4 | ng/ul | 2.127 | 1.100 | 1.93 | 2.23 | DNA | 50.00 |
10 | psb1c3-8 | biospec | 8/1/2015 6:12:51 PM | 90.7 | ng/ul | 1.814 | 0.959 | 1.89 | 1.98 | DNA | 50.00 |
Ligation of „pSB1C3 – Gblocks“
2 | 3 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | ||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Insert | 7.3 ul | 7.0 ul | 4.8 ul | 8.4 ul | 5.8 ul | 8.3 ul | 3.6 ul | 3.9 ul | 3.4 ul | 4.4 ul | 5.9 ul | 3.9 ul | 5.9 ul | |
Vector | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | |
Buffer | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | 2.0 ul | |
Enzyme | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | 1.0 ul | |
ddH₂O | 7.7 ul | 8.0 ul | 10.2 ul | 6.6 ul | 9.2 ul | 6.7 ul | 11.4 ul | 11.1 ul | 11.6 ul | 10.6 ul | 9.1 ul | 11.1 ul | 9.1 ul | |
Total | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 20.0 ul |
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR fwd | VR rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
22X | 55.0 ul | 55.0 ul | 22.0 ul | 22.0 ul | 11.0 ul | 4.4 ul | 271.0 ul | 440.4 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="340"></a>
Result: negative
Colony PCR of „pSB1C3 – Gblocks #14“ (03.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR fwd | VR rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
36X | 90.0 ul | 90.0 ul | 36.0 ul | 36.0 ul | 18.0 ul | 7.2 ul | 442.8 ul | 720.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a> <a href="" data-lightbox="image-1" class="img="left"><img src="" width="360" height="360"></a>
Result: negative
Digestion of “pSB1C3-RFP” (03.08.2015)
Digestion of “pSB1C3-RFP” | ||||||
---|---|---|---|---|---|---|
Vector | Fast Digest Buffer | EcoRI (FD) | PstI (FD) | ddH₂O | Total | |
1x | 2.0 ul | 2.0 ul | 1.0 ul | 1.0 ul | 14.0 ul | 20.0 ul |
37˚C 2h
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Gel Electropheresis was made.
Result: Bands were at the expected section. Gel purification was made.
Ligation of “pSB1C3 – Gblock #14”
Ligation of “pSB1C3 – Gblock #14” | ||||||
---|---|---|---|---|---|---|
Insert | Vector | T4 DNA Ligase Buffer | T4 DNA Ligase | ddH₂O | Total | |
1) PCR clean up vektörü ile 2.5 ul | 2.5 ul | 2.0 ul | ||||
(100ng) | 2.0 ul | |||||
1.0 ul | 12.5 ul | 20.0 ul | ||||
2) Gel Extraction 7.5 ul | 7.5 ul | 0.8 ul | ||||
(50ng) | 2.0 ul | |||||
1.0 ul | 8.7 ul | 20.0 ul |
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/3/2015 6:10:02 PM | 0.2 | ng/ul | 0.004 | -0.010 | -0.41 | 1.13 | DNA | 50.00 |
2 | psb1c3 gel | biospec | 8/3/2015 6:12:16 PM | 62.5 | ng/ul | 1.250 | 0.664 | 1.88 | 0.90 | DNA | 50.00 |
4 | psb1c3 clean-up | biospec | 8/3/2015 6:13:39 PM | 170.5 | ng/ul | 3.409 | 1.884 | 1.81 | 1.33 | DNA | 50.00 |
PCR of “G-Bloks via Q5 Polymerase” (04.08.2015)
PCR | ||||||
---|---|---|---|---|---|---|
SV40 poly A rev | CMV fwd | tetR rev | 2x Q5 Master Mix | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 2.5 ul | 10.0 ul | 5.0 ul | 20.0 ul |
Cycling SV40 rev/CMV fwd | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 63˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1’ | 2’ | 35x |
Cycling CMV fwd/tetR rev | ||||||
---|---|---|---|---|---|---|
98˚C | 98˚C | 66˚C | 72˚C | 72˚C | Cycle | |
Time | 30’’ | 10’’ | 30’’ | 1’ | 2’ | 35x |
Digestion of “pSB1C3-RFP” (04.08.2015)
Digestion | ||||||
---|---|---|---|---|---|---|
Vector | Cut Smart Buffer | EcoRI | SpeI | ddH₂O | Total | |
1x | 2.0 ul | 2.0 ul | 1.0 ul | 1.0 ul | 14.0 ul | 20.0 ul |
37˚C 4h
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Result: Bands were at the expected section.
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/4/2015 7:03:09 AM | 0.9 | ng/ul | 0.017 | 0.005 | 3.41 | 0.68 | DNA | 50.00 |
2 | Blank2 | biospec | 8/4/2015 7:04:51 AM | -0.5 | ng/ul | -0.010 | -0.017 | 0.59 | 0.45 | DNA | 50.00 |
4 | clean up | biospec | 8/4/2015 7:05:55 AM | 86.8 | ng/ul | 1.735 | 0.913 | 1.90 | 0.40 | DNA | 50.00 |
5 | gel ext | biospec | 8/4/2015 7:06:51 AM | 36.2 | ng/ul | 0.723 | 0.380 | 1.90 | 0.29 | DNA | 50.00 |
Colony PCR of “11,12,13 pSB1C3 - GBlocks”
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VF | VR | Tag | ddH₂O | DNA | Total | ||
1x | 2.5 ul | 2.5 ul | 0.5 ul | 0.5 ul | 0.2 ul | 13.3 ul | 5.0 ul | 25.0 ul | |
17X | 42.5 ul | 42.5 ul | 8.5 ul | 8.5 ul | 3.4 ul | 226.1 ul | 340.0 ul |
Cycling CMV fwd/tetR rev | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Result: negative
Ligation and Transformation of “5,6,7,8,9 pSB1C3-GBlocks”
Ligation of “GBlocks 11-12 pSB1C3” | ||||||
---|---|---|---|---|---|---|
Vector | Gen | Buffer | Ligase | ddH₂O | Total | |
5 | 0.8 ul | 2.37 ul | 1.0 ul | 0.5 ul | 5.33 ul | 10.0 ul |
6 | 0.8 ul | 4.2 ul | 1.0 ul | 0.5 ul | 3.5 ul | 10.0 ul |
7 | 0.8 ul | 2.94 ul | 1.0 ul | 0.5 ul | 4.76 ul | 10.0 ul |
8 | 0.8 ul | 4.16 ul | 1.0 ul | 0.5 ul | 3.54 ul | 10.0 ul |
9 | 0.8 ul | 1.79 ul | 1.0 ul | 0.5 ul | 5.91 ul | 10.0 ul |
50ng DNA
1 vector:0.5 insert
Transfomation with TOP10.
Ligation of “GBlocks 11-12 pSB1C3”
Ligation of “GBlocks 11-12 pSB1C3” | ||||||
---|---|---|---|---|---|---|
pSB1C3 | Insert | Buffer | Ligase | ddH₂O | Total | |
11 | 0.8 ul | 1.7 ul | 1.0 ul | 0.5 ul | 6.0 ul | 10.0 ul |
12 | 0.4 ul | 1.1 ul | 1.0 ul | 0.5 ul | 7.0 ul | 10.0 ul |
Transformation was made.
Colony PCR of “5,6,7,8,9,T7 system – Gblocks”
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 0.5 ul | 0.5 ul | 0.5 ul | 0.1 ul | 13.3 ul | 5.0 ul | 24.9 ul |
58X | 145.0 ul | 145.0 ul | 29.0 ul | 29.0 ul | 29.0 ul | 5.8 ul | 771.4 ul | 1154.2 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Ligation of “GBlocks 2,5,6,7,8,9,10,12,13,15,17 pSB1C3-RFP”
Ligation of “GBlocks 11-12 pSB1C3” | ||||||
---|---|---|---|---|---|---|
pSB1C3 | Insert | Buffer | Ligase | ddH₂O | Total | |
2 | 1.4 ul | 3.65 ul | 1.0 ul | 0.5 ul | 3.45 ul | 10.0 ul |
5 | 1.4 ul | 2.4 ul | 1.0 ul | 0.5 ul | 4.7 ul | 10.0 ul |
6 | 1.4 ul | 4.2 ul | 1.0 ul | 0.5 ul | 2.9 ul | 10.0 ul |
7 | 1.4 ul | 2.9 ul | 1.0 ul | 0.5 ul | 4.2 ul | 10.0 ul |
8 | 1.4 ul | 4.15 ul | 1.0 ul | 0.5 ul | 2.95 ul | 10.0 ul |
9 | 1.4 ul | 1.8 ul | 1.0 ul | 0.5 ul | 5.3 ul | 10.0 ul |
10 | 1.4 ul | 1.9 ul | 1.0 ul | 0.5 ul | 5.2 ul | 10.0 ul |
12 | 1.4 ul | 2.2 ul | 1.0 ul | 0.5 ul | 4.9 ul | 10.0 ul |
13 | 1.4 ul | 2.9 ul | 1.0 ul | 0.5 ul | 4.2 ul | 10.0 ul |
15 | 1.4 ul | 2.95 ul | 1.0 ul | 0.5 ul | 4.15 ul | 10.0 ul |
17 | 1.4 ul | 1.25 ul | 1.0 ul | 0.5 ul | 5.75 ul | 10.0 ul |
Transformation was made.
Colony PCR of “pTRE-1,13,15” (06.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | CMV fwd | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
20X | 50.0 ul | 50.0 ul | 20.0 ul | 20.0 ul | 10.0 ul | 2.0 ul | 246.0 ul | 400.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Result: negative
GEL GÖRÜNTÜSÜ
Colony PCR of “5,6,7,9 – Gblocks” (06.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 0.5 ul | 0.5 ul | 0.5 ul | 0.2 ul | 13.3 ul | 5.0 ul | 25.0 ul |
40X | 100.0 ul | 100.0 ul | 20.0 ul | 20.0 ul | 20.0 ul | 8.0 ul | 532.0 ul | 800.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a> <a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Result: negative
Colony PCR of “7,8,10 – Gblocks” from Masterplate
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | T7 terminatör Rev. | T7 terminatör For. | dNTP | Tag | ddH₂O | DNA | Totalzz | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.5 ul | 12.3 ul | 5.0 ul | 25.0 ul |
26X | 65.0 ul | 65.0 ul | 26.0 ul | 26.0 ul | 13.0 ul | 13.0 ul | 319.8 ul | 527.8 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
Digestion of “pET45”
Digestion | |||||||||
---|---|---|---|---|---|---|---|---|---|
pET45 (455ng/ul) | pSB1C3-RFP (400 ng/ul) | Cut Smart Buffer | XhoI (Neb) | BamHI-HF (Neb) | EcoRI-HF (Neb) | SpeI (Neb) | ddH₂O | Total | |
1 | - | 5.0 ul | 2.0 ul | - | - | 1.0 ul | 1.0 ul | 11.0 ul | 20.0 ul |
2 | 4.0 ul | - | 2.0 ul | 1.0 ul | 1.0 ul | - | - | 12.0 ul | 20.0 ul |
37˚C 3.5h
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Result: Positive. Gel extraction was made.
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/9/2015 5:04:14 PM | -0.6 | ng/ul | -0.011 | 0.005 | 3.41 | 0.68 | DNA | 50.00 |
2 | pet45 x+b | biospec | 8/9/2015 5:05:26 PM | 106.9 | ng/ul | 2.138 | -0.017 | 0.59 | 0.45 | DNA | 50.00 |
4 | psb1c3 e+s | biospec | 8/9/2015 5:06:20 PM | 133.3 | ng/ul | 2.666 | 0.913 | 1.90 | 0.40 | DNA | 50.00 |
Ligation | ||||||
---|---|---|---|---|---|---|
Vector | Insert | T4 DNA Ligase Buffer | T4 DNA Ligase | ddH₂O | Total | |
1x | 2.1 ul | 1.0 ul | 2.0 ul | 0.5 ul | 14.4 ul | 20.0 ul |
16˚C 2h
Digestion of “pSB1C3 – HNS”
Ligation | ||||||
---|---|---|---|---|---|---|
Gen | BamHI | XhoI | Cut Smart Buffer | ddH₂O | Total | |
1x | 11.5 ul | 1.0 ul | 1.0 ul | 2.0 ul | 4.5 ul | 20.0 ul |
37˚C 3h
Colony PCR of “pTRE #13”
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | CMV for. | SV40 rev. | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
10X | 25.0 ul | 25.0 ul | 10.0 ul | 10.0 ul | 5.0 ul | 2.0 ul | 123.0 ul | 250.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 60˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="370" height="360"></a>
Result: negative
Colony PCR of “PET45-4,5,6,7,8,9,10”
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | T7 ter. fw | T7 ter. rev | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
86X | 215.0 ul | 215.0 ul | 86.0 ul | 186.0 ul | 43.0 ul | 17.2 ul | 1075.0 ul | 1720.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="https://2015.igem.org/File:ATOMS-Turkiye_11_08_pet45_7,8.jpeg" data-lightbox="image-1" class="img="left"><img src="https://2015.igem.org/File:ATOMS-Turkiye_11_08_pet45_7,8.jpeg" width="410" height="360"></a> <a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a> <a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a> <a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Result: Bands were at the expected section. Liquid Culture was made.
Ligation of “pColA-KanR and pSB1C3-Toehold”
Ligation | |||||||
---|---|---|---|---|---|---|---|
pColA-KanR (500 ng/ul) | pSb1C3-Toehold | T4 DNA Ligase Buffer (Thermo) | T4 DNA Ligase (Thermo) | ddH₂O | Total | Vector:Insert Ratios | |
1 | 2.0 ul | 1.0 ul | 2.0 ul | 1.0 ul | 14.0 ul | 20.0 ul | 1:0.33 |
2 | 2.0 ul | 3.0 ul | 2.0 ul | 1.0 ul | 12.0 ul | 20.0 ul | 1:1 |
22˚C 1h
Mini Prep of “pColA-3,4,5”
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/11/2015 11:11:42 PM | -0.2 | ng/ul | -0.003 | -0.007 | 0.49 | 0.13 | DNA | 50.00 |
2 | ColA-3 | biospec | 8/11/2015 11:13:07 PM | 27.2 | ng/ul | 0.544 | 0.260 | 2.10 | 1.95 | DNA | 50.00 |
3 | ColA-4 | biospec | 8/11/2015 11:14:38 PM | 25.5 | ng/ul | 0.509 | 0.253 | 2.01 | 1.82 | DNA | 50.00 |
4 | ColA-4(2) | biospec | 8/11/2015 11:15:58 PM | 25.8 | ng/ul | 0.517 | 0.255 | 2.022 | 1.81 | DNA | 50.00 |
5 | ColA-5 | biospec | 8/11/2015 11:17:02 PM | 44.4 | ng/ul | 0.887 | 0.449 | 1.98 | 1.76 | DNA | 50.00 |
Result: The Band of pColA were at the expected section. 2000 pb
(CUT CHECK GÖRÜNTÜSÜ)(12.08.2015)
Ligation of “#2 GBlock with pET45” (11.08.2015)
Ligation | ||||||
---|---|---|---|---|---|---|
pET45 (100 ng/ul) | #2 GBlock | T4 DNA Ligase Buffer | T4 DNA Ligase | ddH₂O | Total | |
1x | 1.0 ul | 7.3 ul | 2.0 ul | 1.0 ul | 8.7 ul | 20.0 ul |
22˚C - 1’40’’
Transformation was made.
Mini Prep
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/12/2015 7:38:28 PM | 0.4 | ng/ul | 0.008 | -0.001 | -8.91 | 0.76 | DNA | 50.00 |
2 | pet45 4-4 | biospec | 8/12/2015 7:40:21 PM | 40.3 | ng/ul | 0.807 | 0.402 | 2.01 | 2.44 | DNA | 50.00 |
3 | pet45 4-3 | biospec | 8/12/2015 7:41:10 PM | 53.1 | ng/ul | 1.063 | 0.570 | 1.86 | 1.74 | DNA | 50.00 |
4 | pet45 8-12 | biospec | 8/12/2015 7:41:55 PM | 61.8 | ng/ul | 1.236 | 0.642 | 1.92 | 2.08 | DNA | 50.00 |
5 | pet45 8-3 | biospec | 8/12/2015 7:42:37 PM | 32.9 | ng/ul | 0.658 | 0.332 | 1.98 | 2.00 | DNA | 50.00 |
6 | pet45 9-11 | biospec | 8/12/2015 7:44:03 PM | 34.5 | ng/ul | 0.690 | 0.345 | 2.00 | 1.77 | DNA | 50.00 |
7 | pet45 8-1 | biospec | 8/12/2015 7:44:54 PM | 39.0 | ng/ul | 0.780 | 0.408 | 1.91 | 1.72 | DNA | 50.00 |
8 | pet45 5-10 | biospec | 8/12/2015 7:45:49 PM | 43.2 | ng/ul | 0.865 | 0.441 | 1.96 | 1.81 | DNA | 50.00 |
9 | pet45 6-3 | biospec | 8/12/2015 7:46:42 PM | 43.5 | ng/ul | 0.870 | 0.437 | 1.99 | 1.92 | DNA | 50.00 |
10 | pet45 9-8 | biospec | 8/12/2015 7:47:23 PM | 48.1 | ng/ul | 0.961 | 0.496 | 1.94 | 1.88 | DNA | 50.00 |
11 | pet45 7-4 | biospec | 8/12/2015 7:48:06 PM | 36.2 | ng/ul | 0.723 | 0.372 | 1.94 | 1.97 | DNA | 50.00 |
12 | pet45 4-1 | biospec | 8/12/2015 7:48:50 PM | 49.4 | ng/ul | 0.987 | 0.509 | 1.94 | 1.99 | DNA | 50.00 |
13 | pet45 4-2 | biospec | 8/12/2015 7:49:43 PM | 59.3 | ng/ul | 1.187 | 0.606 | 1.96 | 2.09 | DNA | 50.00 |
14 | pet45 7-3 | biospec | 8/12/2015 7:50:33 PM | 30.2 | ng/ul | 0.604 | 0.307 | 1.97 | 1.96 | DNA | 50.00 |
15 | psb1c3 12 | biospec | 8/12/2015 7:51:18 PM | 141.1 | ng/ul | 2.823 | 1.462 | 1.93 | 2.14 | DNA | 50.00 |
16 | pet45 5-7 | biospec | 8/12/2015 7:52:04 PM | 26.1 | ng/ul | 0.522 | 0.265 | 1.97 | 2.00 | DNA | 50.00 |
17 | pet45 5-3 | biospec | 8/12/2015 7:52:49 PM | 46.1 | ng/ul | 0.923 | 0.470 | 1.96 | 2.02 | DNA | 50.00 |
18 | pet45 6-12 | biospec | 8/12/2015 7:53:36 PM | 29.3 | ng/ul | 0.586 | 0.283 | 2.07 | 1.80 | DNA | 50.00 |
19 | pet45 1-12 | biospec | 8/12/2015 7:54:25 PM | 21.4 | ng/ul | 0.428 | 0.210 | 2.04 | 1.72 | DNA | 50.00 |
20 | pet45 5-8 | biospec | 8/12/2015 7:55:08 PM | 29.1 | ng/ul | 0.582 | 0.299 | 1.95 | 2.07 | DNA | 50.00 |
21 | psb1c3 1 | biospec | 8/12/2015 7:55:51 PM | 75.3 | ng/ul | 1.507 | 0.779 | 1.93 | 2.07 | DNA | 50.00 |
22 | pet45 8-4 | biospec | 8/12/2015 7:56:33 PM | 44.1 | ng/ul | 0.882 | 0.451 | 1.96 | 1.99 | DNA | 50.00 |
23 | pet45 7-2 | biospec | 8/12/2015 7:57:16 PM | 25.3 | ng/ul | 0.506 | 0.259 | 1.95 | 1.70 | DNA | 50.00 |
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/12/2015 7:17:01 PM | -0.3 | ng/ul | -0.007 | -0.015 | 0.47 | 0.23 | DNA | 50.00 |
2 | psb1c3-4hns | biospec | 8/12/2015 7:18:16 PM | 61.9 | ng/ul | 1.237 | 0.652 | 1.90 | 2.17 | DNA | 50.00 |
3 | pet45 10-1 | biospec | 8/12/2015 7:19:32 PM | 46.7 | ng/ul | 0.934 | 0.486 | 1.92 | 2.00 | DNA | 50.00 |
4 | pet45 10-6 | biospec | 8/12/2015 7:20:23 PM | 52.0 | ng/ul | 1.040 | 0.549 | 1.89 | 1.88 | DNA | 50.00 |
5 | pet45 10-3 | biospec | 8/12/2015 7:21:14 PM | 51.9 | ng/ul | 1.037 | 0.536 | 1.94 | 2.10 | DNA | 50.00 |
6 | pet45 9-6 | biospec | 8/12/2015 7:22:10 PM | 37.7 | ng/ul | 0.755 | 0.389 | 1.94 | 1.99 | DNA | 50.00 |
7 | pet45 10-8 | biospec | 8/12/2015 7:23:08 PM | 85.3 | ng/ul | 1.705 | 1.002 | 1.70 | 0.93 | DNA | 50.00 |
8 | psb1c3 16-trigger rna | biospec | 8/12/2015 7:24:27 PM | 125.7 | ng/ul | 2.514 | 1.401 | 1.79 | 1.32 | DNA | 50.00 |
9 | pet45 10 | biospec | 8/12/2015 7:25:23 PM | 84.7 | ng/ul | 1.694 | 1.025 | 1.65 | 0.76 | DNA | 50.00 |
10 | pet45 6-6 | biospec | 8/12/2015 7:26:23 PM | 100.3 | ng/ul | 2.006 | 1.124 | 1.79 | 1.24 | DNA | 50.00 |
11 | pet45 8-2 | biospec | 8/12/2015 7:27:20 PM | 67.5 | ng/ul | 1.349 | 0.778 | 1.73 | 0.91 | DNA | 50.00 |
12 | pet45 7-1 | biospec | 8/12/2015 7:28:07 PM | 52.5 | ng/ul | 1.050 | 0.549 | 1.91 | 1.83 | DNA | 50.00 |
Cut Check of “pET45 1,4,5,6,7,8,9,10 – BamHI/XhoI”
Ligation | ||||||
---|---|---|---|---|---|---|
Gen (250 ng/ul) | Cut Smart Buffer | BamHI (NEB) | XhoI (NEB) | ddH₂O | Total | |
1 | 11.7 ul | 2.0 ul | 1.0 ul | 1.0 ul | 4.3 ul | 20.0 ul |
4 | 6.2 ul | 2.0 ul | 1.0 ul | 1.0 ul | 9.8 ul | 20.0 ul |
5 | 8.6 ul | 2.0 ul | 1.0 ul | 1.0 ul | 7.4 ul | 20.0 ul |
6 | 8.6 ul | 2.0 ul | 1.0 ul | 1.0 ul | 7.4 ul | 20.0 ul |
7 | 8.3 ul | 2.0 ul | 1.0 ul | 1.0 ul | 7.7 ul | 20.0 ul |
8 | 7.4 ul | 2.0 ul | 1.0 ul | 1.0 ul | 8.6 ul | 20.0 ul |
9 | 6.7 ul | 2.0 ul | 1.0 ul | 1.0 ul | 9.3 ul | 20.0 ul |
10 | 5.9 ul | 2.0 ul | 1.0 ul | 1.0 ul | 10.1 ul | 20.0 ul |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a> <a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/13/2015 1:31:59 AM | 0.0 | ng/ul | -0.001 | -0.005 | 0.17 | 0.08 | DNA | 50.00 |
2 | psb1c3-e+p | biospec | 8/13/2015 1:32:58 AM | 58.6 | ng/ul | 1.172 | 0.618 | 1.90 | 0.57 | DNA | 50.00 |
Colony PCR of “pTRE – 12,13 GBlocks”
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | CMV fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
15X | 37.5 ul | 37.5 ul | 15.0 ul | 15.0 ul | 7.5 ul | 3.0 ul | 184.5 ul | 300.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="https://2015.igem.org/File:ATOMS-Turkiye_14_08_ptre_12,13.jpeg" data-lightbox="image-1" class="img="left"><img src="https://2015.igem.org/File:ATOMS-Turkiye_14_08_ptre_12,13.jpeg" width="410" height="360"></a>
Result: negative
Colony PCR of “pColA – Toehold GFP”
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pColA fw | pColA rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
26X | 65.0 ul | 65.0 ul | 26.0 ul | 26.0 ul | 13.0 ul | 5.2 ul | 319.8 ul | 520.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Result: 2-4, 2-8, 1-6 Bands were at the expected section.
Digestion of “pSB1C3-RFP”
Digestion | ||||||
---|---|---|---|---|---|---|
pSB1C3-RFP (400 ng/ul) | Cut Smart Buffer | EcoRI-HF | PstI | ddH₂O | Total | |
1 | 5.0 ul | 2.0 ul | 1.0 ul | 1.0 ul | 11.0 ul | 20.0 ul |
2 | 5.0 ul | 2.0 ul | 1.0 ul | 1.0 ul | 11.0 ul | 20.0 ul |
3 | 5.0 ul | 2.0 ul | 1.0 ul | 1.0 ul | 11.0 ul | 20.0 ul |
37˚C – 3.5h
Gel extraction was made.
Ligation of “pSB1C3-GBlocks”
Ligation | ||||||
---|---|---|---|---|---|---|
Digested pSB1C3 | GBlocks | T4 DNA Ligase Buffer | T4 DNA Ligase | ddH₂O | Total | |
1 | 1.7 ul | 2.0 ul | 2.0 ul | 1.0 ul | 13.3 ul | 20.0 ul |
22˚C – 1.30h
Transformation with Neb10-β. Colony PCR will be made.
Colony PCR of “GBlocks – 2,5,6,7,8,9” (14.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
45X | 112.5 ul | 112.5 ul | 45.0 ul | 45.0 ul | 22.5 ul | 9.0 ul | 553.5 ul | 900.0 ul | |
55X | 137.5 ul | 137.5 ul | 55.0 ul | 55.0 ul | 27.5 ul | 11.0 ul | 676.5 ul | 1100.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
<a href="" data-lightbox="image-1" class="img="left"><img src="" width="410" height="360"></a>
Digestion/Gel Electrophoresis/Gel Extraction and Purification of “pSB1C3”
Digestion | ||||||
---|---|---|---|---|---|---|
DNA | FD | EcoRI | PstI | ddH₂O | Total | |
1x | 2.5 ul | 2.0 ul | 1.0 ul | 1.0 ul | 13.5 ul | 20.0 ul |
37˚C 1h
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/16/2015 10:45:30 PM | 0.1 | ng/ul | 0.003 | -0.017 | 0.17 | -0.14 | DNA | 50.00 |
2 | psb1c3-e+p | biospec | 8/16/2015 10:47:18 PM | 14.0 | ng/ul | 0.279 | 0.145 | 1.92 | 0.15 | DNA | 50.00 |
Colony PCR of “pSB1C3 7,10,12,13 and pET45 2” (14.08.2015)
Colony PCR of “pSB1C3 7,10,12,13” | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
6X | 15.0 ul | 15.0 ul | 6.0 ul | 6.0 ul | 3.0 ul | 1.2 ul | 73.8 ul | 120.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
Colony PCR of “pET45 2” | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
12X | 30.0 ul | 30.0 ul | 12.0 ul | 12.0 ul | 6.0 ul | 2.4 ul | 147.6 ul | 240.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/16/2015 8:54:05 PM | -0.2 | ng/ul | -0.004 | -0.017 | 0.25 | 0.12 | DNA | 50.00 |
2 | psb1c3 13-15 | biospec | 8/16/2015 8:55:29 PM | 125.2 | ng/ul | 2.505 | 1.339 | 1.87 | 2.01 | DNA | 50.00 |
3 | psb1c3 15-19 | biospec | 8/16/2015 8:56:24 PM | 93.5 | ng/ul | 1.871 | 0.999 | 1.87 | 1.94 | DNA | 50.00 |
4 | psb1c3 7-16 | biospec | 8/16/2015 8:57:33 PM | 149.4 | ng/ul | 2.988 | 1.575 | 1.90 | 2.14 | DNA | 50.00 |
5 | psb1c3 10-16 | biospec | 8/16/2015 8:58:18 PM | 138.0 | ng/ul | 2.760 | 1.469 | 1.88 | 2.13 | DNA | 50.00 |
6 | psb1c3 10-17 | biospec | 8/16/2015 8:59:03 PM | 185.5 | ng/ul | 3.710 | 1.994 | 1.86 | 1.86 | DNA | 50.00 |
7 | psb1c3 7-11 | biospec | 8/16/2015 8:00:54 PM | 73.8 | ng/ul | 1.475 | 0.843 | 1.75 | 1.08 | DNA | 50.00 |
8 | psb1c3 13-14 | biospec | 8/16/2015 8:01:53 PM | 178.7 | ng/ul | 3.575 | 1.922 | 1.86 | 1.87 | DNA | 50.00 |
9 | psb1c3 7-20 | biospec | 8/16/2015 8:02:54 PM | 73.6 | ng/ul | 1.472 | 0.774 | 1.90 | 2.05 | DNA | 50.00 |
10 | pcola toe 2-4 | biospec | 8/16/2015 8:03:57 PM | 40.1 | ng/ul | 0.801 | 0.441 | 1.82 | 1.13 | DNA | 50.00 |
11 | pcola toe 2-8 | biospec | 8/16/2015 8:05:31 PM | 48.7 | ng/ul | 0.974 | 0.503 | 1.94 | 1.61 | DNA | 50.00 |
12 | pcola toe 1-6 | biospec | 8/16/2015 8:06:09 PM | 54.9 | ng/ul | 1.098 | 0.570 | 1.93 | 1.62 | DNA | 50.00 |
13 | pcola blunt 1 | biospec | 8/16/2015 8:06:52 PM | 23.9 | ng/ul | 0.479 | 0.244 | 1.96 | 1.24 | DNA | 50.00 |
14 | pcola blunt 6 | biospec | 8/16/2015 8:07:30 PM | 31.1 | ng/ul | 0.623 | 0.314 | 1.98 | 1.67 | DNA | 50.00 |
Ligation | |||||||
---|---|---|---|---|---|---|---|
pColA (Vector 50 ng/ul) | Insert (Damp-Pex) | T4 DNA Ligase Buffer | T4 DNA Ligase | ddH₂O | Total | Vector:Insert Ratio | |
9.1 | 2.0 ul | 1.0 ul | 2.0 ul | 1.0 ul | 14.0 ul | 20.0 ul | 1:0.33 |
9.2 | 2.0 ul | 2.8 ul | 2.0 ul | 1.0 ul | 12.2 ul | 20.0 ul | 1:1 |
Miniprep of “pTet-off DH5α /Top10/NEB5fα/NEB10β”
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/18/2015 3:04:27 PM | 0.4 | ng/ul | 0.008 | -0.008 | -1.01 | 1.01 | DNA | 50.00 |
2 | top10 | biospec | 8/18/2015 3:05:19 PM | 258.5 | ng/ul | 5.169 | 2.753 | 1.88 | 2.11 | DNA | 50.00 |
3 | neb5a | biospec | 8/18/2015 3:06:25 PM | 30.6 | ng/ul | 0.612 | 0.324 | 1.89 | 1.34 | DNA | 50.00 |
4 | neb5a | biospec | 8/18/2015 3:07:30 PM | 33.4 | ng/ul | 0.668 | 0.350 | 1.91 | 1.27 | DNA | 50.00 |
5 | neb10beta | biospec | 8/18/2015 3:08:22 PM | 214.6 | ng/ul | 4.292 | 2.264 | 1.90 | 2.19 | DNA | 50.00 |
Colony PCR of “pET45 #2.8” (15.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
25X | 62.5 ul | 62.5 ul | 25.0 ul | 25.0 ul | 12.5 ul | 5.0 ul | 307.5 ul | 500.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
Colony PCR of “pSB1C3 17,6,8” (S. 118/15.08.2015)
Colony PCR of “pSB1C3 17,6,8” | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
32X | 80.0 ul | 80.0 ul | 32.0 ul | 32.0 ul | 16.0 ul | 6.4 ul | 393.6 ul | 640.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: 6: Liquid Culture was made.
8: sequencing will be done
17: Colony PCR will be repeated.
GEL GÖRÜNTÜSÜ
Colony PCR of “pColA blunt”
Colony PCR of “pColA blunt” | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | ColA fw | ColA rev | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
20X | 50.0 ul | 50.0 ul | 20.0 ul | 20.0 ul | 10.0 ul | 4.0 ul | 246.0 ul | 400.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
PCR of “Q5 DNA Polymerase for pCAG” (19.08.2015)
PCR | ||||||
---|---|---|---|---|---|---|
Template DNA | pCAG fw | pCAG rev | Master 2x Mix | ddH₂O | Total | |
1x | 1.0 ul | 5.0 ul | 5.0 ul | 25.0 ul | 14.0 ul | 50.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
98˚C | 98˚C | 70˚C | 72˚C | 72˚C | Cycle | |
Time | 30’’ | 10’’ | 30’’ | 1’ | 5’ | 35x |
Result: Bands were at the expected section. PCR clean up will be made.
GEL GÖRÜNTÜSÜ
Digestion of “pCAG – Clean up”
Digestion | |||||||
---|---|---|---|---|---|---|---|
pCAG – Clean up (34 ng/ul) | pCAG – Clean up (61.6 ng/ul) | EcoRI-HF (NEB) | XhoI (NEB) | Cut Smart Buffer | ddH₂O | Total | |
1 | 7.4 ul | - | 0.5 ul | 0.5 ul | 2.0 ul | 9.6 ul | 20.0 ul |
2 | - | 4.1 ul | 0.5 ul | 0.5 ul | 2.0 ul | 12.9 ul | 20.0 ul |
37˚C overnight
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/19/2015 7:08:39 PM | 0.0 | ng/ul | 0.000 | 0.000 | 27.59 | -0.07 | DNA | 50.00 |
2 | pcag | biospec | 8/19/2015 7:10:01 PM | 61.6 | ng/ul | 1.233 | 0.686 | 1.80 | 1.02 | DNA | 50.00 |
Digestion of “pSB1C3 – mLacI miR373-BS” (19.08.2015)
Digestion | |||||||
---|---|---|---|---|---|---|---|
mLacI miR373-BS | EcoRI-HF (NEB) | XhoI (NEB) | Cut Smart Buffer | ddH₂O | Total | ||
1 | 3.4 ul | 0.5 ul | 0.5 ul | 2.0 ul | 13.9 ul | 20.0 ul |
37˚C 2.30h
Ligation of “pSB1C3 – mLacI miR373-BS with pTRE”
Ligation | |||||||
---|---|---|---|---|---|---|---|
Digested mLacI miR373-BS with pTRE | pTRE | T4 DNA Ligase Buffer | T4 DNA Ligase | Cut Smart Buffer | ddH₂O | Total | |
1 | 3.0 ul | 1.0 ul | 2.0 ul | 1.0 ul | 2.0 ul | 13.0 ul | 20.0 ul |
22˚C 1.30h
Transformation will be made.
Colony PCR of “#6,8” (20.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | ColA fw | ColA rev | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
25X | 62.5 ul | 62.5 ul | 25.0 ul | 25.0 ul | 12.5 ul | 5.0 ul | 307.5 ul | 500.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: Positive. Liquid Culture was made.
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/22/2015 12:11:34 PM | 0.5 | ng/ul | 0.010 | -0.003 | -3.39 | -3.90 | DNA | 50.00 |
2 | 8-1 | biospec | 8/22/2015 12:12:38 PM | 70.3 | ng/ul | 1.406 | 0.755 | 1.86 | 1.64 | DNA | 50.00 |
3 | 8-3 | biospec | 8/22/2015 12:13:38 PM | 80.7 | ng/ul | 1.613 | 0.846 | 1.91 | 2.11 | DNA | 50.00 |
4 | 6-1 | biospec | 8/22/2015 12:14:32 PM | 33.3 | ng/ul | 0.666 | 0.354 | 1.88 | 1.44 | DNA | 50.00 |
Cut Check of “#6,8”
Cut Check | ||||||
---|---|---|---|---|---|---|
DNA (400 ng/ul) | EcoRI | PstI | Fast Digest Buffer | ddH₂O | Total | |
1 | 5.7 ul | 0.5 ul | 0.5 ul | 2.0 ul | 11.3 ul | 20.0 ul |
2 | 4.8 ul | 0.5 ul | 0.5 ul | 2.0 ul | 12.2 ul | 20.0 ul |
3 | 12.0 ul | 0.5 ul | 0.5 ul | 2.0 ul | 5.0 ul | 20.0 ul |
37˚C 40’
Result: positive
GEL GÖRÜNTÜSÜ
Colony PCR “pSB1C3 17,6,8 – pColA, Damp-Pex, pTetoff” (19.08.2015)
GEL GÖRÜNTÜLERI
Mini Prep of “pSB1C3-#17”
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/20/2015 5:04:31 PM | 0.6 | ng/ul | 0.013 | 0.007 | 1.93 | 1.40 | DNA | 50.00 |
2 | psb1c3 17-2 | biospec | 8/20/2015 5:05:28 PM | 78.9 | ng/ul | 1.579 | 0.821 | 1.92 | 2.17 | DNA | 50.00 |
3 | psb1c3 17-5 | biospec | 8/20/2015 5:06:18 PM | 77.5 | ng/ul | 1.551 | 0.806 | 1.92 | 2.08 | DNA | 50.00 |
4 | psb1c3 17-8 | biospec | 8/20/2015 5:07:02 PM | 87.5 | ng/ul | 1.749 | 0.910 | 1.92 | 2.08 | DNA | 50.00 |
5 | psb1c3 17-10 | biospec | 8/20/2015 5:07:47 PM | 105.4 | ng/ul | 2.108 | 1.087 | 1.94 | 2.15 | DNA | 50.00 |
Digestion of “pSB1C3 T7-10 and pET45 #4-1, #5-3, #6-6, #8-15” (21.08.2015)
Digestion pET45 #4-1, #5-3, #6-6, #8-15 | |||||||
---|---|---|---|---|---|---|---|
DNA | BamHI-HF (NEB) | XhoI (NEB) | Cut Smart Buffer | ddH₂O | Total | ||
#4-1 | 10.1 ul | 1.0 ul | 1.0 ul | 2.0 ul | 5.9 ul | 20.0 ul | |
#5-3 | 10.8 ul | 1.0 ul | 1.0 ul | 2.0 ul | 5.2 ul | 20.0 ul | |
#6-6 | 5.0 ul | 1.0 ul | 1.0 ul | 2.0 ul | 11.0 ul | 20.0 ul | |
#8-15 | 16.0 ul | 1.0 ul | 1.0 ul | 2.0 ul | - | 20.0 ul |
37˚C 3h
Digestion pSB1C3 T7-10 | |||||||
---|---|---|---|---|---|---|---|
DNA | BamHI-HF (NEB) | XhoI (NEB) | Cut Smart Buffer | ddH₂O | Total | ||
#4-1 | 4.8 ul | 1.0 ul | 1.0 ul | 2.0 ul | 11.2 ul | 20.0 ul |
1ul rSAP / 37˚C 1h
Ligation | |||||||
---|---|---|---|---|---|---|---|
Vector (pSB1C3) | Insert | T4 DNA Ligase Buffer | T4 DNA Ligase | ddH₂O | Total | ||
#4 | 1.0 ul | 1.0 ul | 2.0 ul | 1.0 ul | 15.0 ul | 20.0 ul | |
#5 | 1.0 ul | 1.0 ul | 2.0 ul | 1.0 ul | 15.0 ul | 20.0 ul | |
#6 | 1.0 ul | 1.0 ul | 2.0 ul | 1.0 ul | 15.0 ul | 20.0 ul | |
#8 | 1.0 ul | 1.0 ul | 2.0 ul | 1.0 ul | 15.0 ul | 20.0 ul |
22˚C 2h
-80 ˚C 20’’ Heat Inaktivation
Colony PCR of “pTRE - #13 mLacI-miR373 BS” (21.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | CMV fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
22X | 55.0 ul | 55.0 ul | 22.0 ul | 22.0 ul | 11.0 ul | 4.4 ul | 270.6 ul | 440.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Results: negative
GEL GÖRÜNTÜLERI
Mini Prep of “pSB1C3 #6-25,26”
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/20/2015 10:01:33 AM | 0.4 | ng/ul | 0.007 | -0.003 | -2.12 | -1.13 | DNA | 50.00 |
2 | psb1c3 #6-25 | biospec | 8/20/2015 10:02:45 AM | 41.0 | ng/ul | 0.820 | 0.421 | 1.95 | 1.93 | DNA | 50.00 |
3 | psb1c3 #6-26 | biospec | 8/20/2015 10:03:50 AM | 46.0 | ng/ul | 0.920 | 0.464 | 1.98 | 1.97 | DNA | 50.00 |
Cut Check of “pSB1C3 #6-25,26”
Cut Check | ||||||
---|---|---|---|---|---|---|
DNA | EcoRI-HF | PstI-HF | Cut Smart Buffer | ddH₂O | Total | |
#6-25 | 6.1 ul | 0.5 ul | 0.5 ul | 2.0 ul | 10.9 ul | 20.0 ul |
#6-26 | 5.5 ul | 0.5 ul | 0.5 ul | 2.0 ul | 11.5 ul | 20.0 ul |
37˚C 3h
Result: negative
Digestion of “pTRE #12”
Digestion | ||||||
---|---|---|---|---|---|---|
pTRE #12 (5000 ng/ul) | EcoRI-HF | XhoI (NEB) | Cut Smart Buffer | ddH₂O | Total | |
1x | 16.0 ul | 0.5 ul | 0.5 ul | 2.0 ul | 1.0 ul | 20.0 ul |
37˚C 3h
Result: negative
GEL GÖRÜNTÜSÜ
Digestion of “Damp-Pex pSB1C3” (22.08.2015)
Digestion pSB1C3 T7-10 | |||||||
---|---|---|---|---|---|---|---|
DNA (500 ng/ul) | NotI | Cut Smart Buffer | ddH₂O | Total | |||
#9-5 | 3.42 ul | 1.0 ul | 2.0 ul | 13.58 ul | 20.0 ul | ||
#9-10 | 3.56 ul | 1.0 ul | 2.0 ul | 13.44 ul | 20.0 ul |
37˚C 3h
Ligation of “Damp-Pex pColA”
Ligation | |||||||
---|---|---|---|---|---|---|---|
pColA (100 ng/ul) | Insert (36 ng/ul) | T4 DNA Ligase Buffer | T4 DNA Ligase | ddH₂O | Total | ||
1x | 2.8 ul | 5.41 ul | 2.0 ul | 1.0 ul | 15.0 ul | 20.0 ul |
22˚C 1.30h
Transformation with NEB5α.
Colony PCR of “Damp-Pex + pColA” (23.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pColA fw | pColA rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
26X | 65.0 ul | 65.0 ul | 26.0 ul | 26.0 ul | 13.0 ul | 5.2 ul | 319.8 ul | 520.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: #5-4, #5-9 Liquid Culture was made.
GEL GÖRÜNTÜSÜ
Mini Prep of “pTetoff #14-4,5,8” (22.08.2015)
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/22/2015 11:01:09 AM | 0.1 | ng/ul | 0.002 | -0.008 | -0.23 | -0.10 | DNA | 50.00 |
2 | ptetoff 14-4 | biospec | 8/22/2015 11:02:05 AM | 159.6 | ng/ul | 3.193 | 1.678 | 1.90 | 2.00 | DNA | 50.00 |
3 | ptetoff 14-5 | biospec | 8/22/2015 11:03:21 AM | 141.1 | ng/ul | 2.822 | 1.466 | 1.93 | 1.97 | DNA | 50.00 |
4 | ptetoff 14-8 | biospec | 8/22/2015 11:04:14 AM | 367.5 | ng/ul | 7.350 | 3.969 | 1.85 | 2.11 | DNA | 50.00 |
Cut Check I | ||||||
---|---|---|---|---|---|---|
DNA (250 ng/ul) | SalI (FD) | HindIII (FD) | Fast Digest Buffer | ddH₂O | Total | |
#14-4 | 1.6 ul | 0.5 ul | 0.5 ul | 2.0 ul | 15.4 ul | 20.0 ul |
#14-5 | 1.8 ul | 0.5 ul | 0.5 ul | 2.0 ul | 15.2 ul | 20.0 ul |
#14-8 | 0.7 ul | 0.5 ul | 0.5 ul | 2.0 ul | 16.3 ul | 20.0 ul |
37˚C 2h
Result: negative
GEL GÖRÜNTÜSÜ
Cut Check II | ||||||
---|---|---|---|---|---|---|
DNA (250 ng/ul) | SalI (FD) | NotI (FD) | Fast Digest Buffer | ddH₂O | Total | |
#14-4 | 1.6 ul | 0.5 ul | 0.5 ul | 2.0 ul | 15.4 ul | 20.0 ul |
#14-5 | 1.8 ul | 0.5 ul | 0.5 ul | 2.0 ul | 15.2 ul | 20.0 ul |
#14-8 | 0.7 ul | 0.5 ul | 0.5 ul | 2.0 ul | 16.3 ul | 20.0 ul |
37˚C 30’’
Result: negative
GEL GÖRÜNTÜSÜ
Colony PCR of “pSB1C3 – T7 #4,5,6,8” (23.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
50x | 125.0 ul | 125.0 ul | 50.0 ul | 50.0 ul | 25.0 ul | 10.0 ul | 615.0 ul | 1000.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
GEL GÖRÜNTÜSÜ </div> </section>
<section class="accordion">
<input id"august-week4" name="august-week4" type="checkbox" /> <label for="august-week4">Week 1, 24.08.-31.08.</label> <article class="ac-small">
</html> Digestion of “pTRE #12-2” (24.08.2015)
Digestion | |||||||
---|---|---|---|---|---|---|---|
pTRE #12-2 (2000 ng/ul) | EcoRI-HF | XhoI-NEB | Cut Smart Buffer | ddH₂O | Total | ||
1x | 6.4 ul | 0.5 ul | 0.5 ul | 2.0 ul | 10.6 ul | 20.0 ul |
37˚C 2h
Result: Bands were at the expected section. Gel Extraction was made.
Mini Prep
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/24/2015 1:28:49 AM | 0.0 | ng/ul | 0.001 | -0.013 | -0.07 | -0.07 | DNA | 50.00 |
2 | ptre gel | biospec | 8/24/2015 1:30:29 AM | 15.3 | ng/ul | 0.306 | 0.164 | 1.87 | 0.65 | DNA | 50.00 |
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/24/2015 1:26:27 AM | 0.7 | ng/ul | 0.015 | 0.001 | 18.47 | 0.53 | DNA | 50.00 |
2 | ptre | biospec | 8/24/2015 1:29:38 AM | 13.7 | ng/ul | 0.275 | 0.145 | 1.90 | 0.10 | DNA | 50.00 |
Digestion of “TEV-Protease pET45/pSB1C3 T7/pColA (24.08.2015)
Digestion | |||||||
---|---|---|---|---|---|---|---|
DNA (500 ng/ul) | BamHI-HF | XhoI-HF | Cut Smart Buffer | ddH₂O | Total | ||
1x | 5.86 ul | 0.5 ul | 0.5 ul | 2.0 ul | 11.14 ul | 20.0 ul |
37˚C 3h
Ligation of “pCAGop with pTRE #12-2” (24.08.2015)
Ligation | |||||||
---|---|---|---|---|---|---|---|
pCAG (18.38 ng/ul) | pTRE #12-2 (100 ng/ul) | T4 DNA Ligase | T4 DNA Ligase Buffer | Cut Smart Buffer | ddH₂O | Total | |
1x | 0.6 ul | 6.6 ul | 1.0 ul | 2.0 ul | 2.0 ul | 9.8 ul | 20.0 ul |
Transformation was made.
Cut Check “pSB1C3-T7 #4,5,6,8” (24.08.2015)
Cut Check | ||||||
---|---|---|---|---|---|---|
DNA (300 ng/ul) | EcoRI (FD) | PstI (FD) | Fast Digest Buffer | ddH₂O | Total | |
1x | 3.5 ul | 1.0 ul | 1.0 ul | 2.0 ul | 12.5 ul | 20.0 ul |
30’ digestion
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/25/2015 1:26:12 PM | 0.8 | ng/ul | 0.016 | -0.003 | -5.85 | 2.25 | DNA | 50.00 |
2 | #9-5-4 | biospec | 8/25/2015 1:27:29 PM | 59.1 | ng/ul | 1.182 | 0.603 | 1.96 | 2.25 | DNA | 50.00 |
3 | #9-5-9 | biospec | 8/25/2015 1:28:30 PM | 37.9 | ng/ul | 0.758 | 0.375 | 2.02 | 2.19 | DNA | 50.00 |
Colony PCR of “pTRE #13” (25.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | CMV fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
7x | 17.5 ul | 17.5 ul | 7.0 ul | 7.0 ul | 3.5 ul | 1.4 ul | 86.1 ul | 140.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
GEL GÖRÜNTÜSÜ + pTRE13 cut check görüntüsü
Colony PCR of “pCAG-DsRed” (25.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
14x | 35.0 ul | 35.0 ul | 14.0 ul | 14.0 ul | 7.0 ul | 2.8 ul | 172.2 ul | 280.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: Bands weren’t at the expected section.
GEL GÖRÜNTÜSÜ
Cut Check of „pSB1C3 – TlpB“ (25.05.2015)
Cut Check | ||||||
---|---|---|---|---|---|---|
DNA (250 ng/ul) | EcoRI (FD) | Fast Digest Buffer | ddH₂O | Total | ||
#8-1 | 3.6 ul | 0.5 ul | 2.0 ul | 13.9 ul | 20.0 ul | |
#8-3 | 3.1 ul | 0.5 ul | 2.0 ul | 14.4 ul | 20.0 ul |
7˚C 35’
The colonies will be made glycerol stock.
GEL GÖRÜNTÜSÜ
Ligation of “Damp-Pex” (25.08.2015)
Ligation | |||||||
---|---|---|---|---|---|---|---|
DNA (500 ng/ul) | NotI | Cut Smart Buffer | ddH₂O | Total | |||
#9-5 | 3.78 ul | 1.0 ul | 2.0 ul | 13.22 ul | 20.0 ul | ||
#9-10 | 3.87 ul | 1.0 ul | 2.0 ul | 13.13 ul | 20.0 ul |
37˚C 3h
Ligation | |||||||
---|---|---|---|---|---|---|---|
ColA (100 ng/ul) | Insert (36 ng/ul) | T4 DNA Ligase | T4 DNA Ligase Buffer | ddH₂O | Total | ||
#9-5 | 10.0 ul | 14.4 ul | 1.0 ul | 3.0 ul | 1.6 ul | 30.0 ul |
22˚C 1.5h
Transformation was made.
Colony PCR of “pCAGop-DsRed” (26.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
10x | 25.0 ul | 25.0 ul | 10.0 ul | 10.0 ul | 5.0 ul | 2.0 ul | 123.0 ul | 200.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: 2 colonie will be made liquid culture and streak plate. Ligation and transformation will be repeated.
GEL GÖRÜNTÜSÜ
Mini Prep of “pTRE #13-3,5” (26.08.2015)
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/26/2015 4:55:28 PM | 0.1 | ng/ul | 0.003 | -0.020 | -0.13 | -0.13 | DNA | 50.00 |
2 | ptre 13-3 | biospec | 8/26/2015 4:56:47 PM | 77.9 | ng/ul | 1.558 | 0.772 | 2.02 | 1.67 | DNA | 50.00 |
3 | ptre 13-5 | biospec | 8/26/2015 4:58:06 PM | 76.0 | ng/ul | 1.520 | 0.776 | 1.96 | 1.48 | DNA | 50.00 |
Colony PCR of “pColA – Damp-Pex, pSB1C3 – TEV
Colony PCR “pColA – Damp-Pex” | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
10x | 25.0 ul | 25.0 ul | 10.0 ul | 10.0 ul | 5.0 ul | 2.0 ul | 123.0 ul | 200.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Colony PCR “pSB1C3 – TEV Protease”
Colony PCR “pSB1C3 – TEV Protease” | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
10x | 25.0 ul | 25.0 ul | 10.0 ul | 10.0 ul | 5.0 ul | 2.0 ul | 123.0 ul | 200.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
Colony PCR “pSB1C3 – TEV Protease” – Repeat (27.08.2015)
Colony PCR “pSB1C3 – TEV Protease” | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VR | VF | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
15x | 37.5 ul | 37.5 ul | 15.0 ul | 15.0 ul | 7.5 ul | 3.0 ul | 184.5 ul | 300.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
Colony PCR of “pTRE – Luc #14” (27.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | pTRE Luc rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
14x | 35.0 ul | 35.0 ul | 14.0 ul | 14.0 ul | 7.0 ul | 2.8 ul | 172.2 ul | 280.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: Bands weren’t at the expected section.
GEL GÖRÜNTÜSÜ
Gel Extraction of “Damp-Pex”
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/27/2015 9:39:17 PM | 1.1 | ng/ul | 0.022 | 0.009 | 2.36 | 1.00 | DNA | 50.00 |
2 | damp-pex | biospec | 8/27/2015 9:40:04 PM | 8.2 | ng/ul | 0.164 | 0.090 | 1.82 | 0.05 | DNA | 50.00 |
Colony PCR of “pCAGop-DsRed #12” (27.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
14x | 35.0 ul | 35.0 ul | 14.0 ul | 14.0 ul | 7.0 ul | 2.8 ul | 172.2 ul | 280.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: Negative.
GEL GÖRÜNTÜSÜ
Ligation of “pCAGop-DsRed” (27.08.2015)
Ligation | ||||||
---|---|---|---|---|---|---|
pCAG (18.38 ng/ul) | pTRE #12 (100 ng/ul) | T4 DNA Ligase | T4 DNA Ligase Buffer | ddH₂O | Total | |
1x | 0.3 ul | 6.6 ul | 1.0 ul | 2.0 ul | 10.1 ul | 20.0 ul |
22˚C 1.30h
Transformation was made.
Colony PCR of „pSB1C3-T7 #5“ (27.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
8x | 20.0 ul | 20.0 ul | 8.0 ul | 8.0 ul | 4.0 ul | 1.6 ul | 98.4 ul | 160.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: #1-1 Liquid Culture was made.
GEL GÖRÜNTÜSÜ
Digestion of “pSB1C3-T7 TEV Protease (17. and 18. Colony)” (27.08.2015)
Digestion | |||||
---|---|---|---|---|---|
DNA (500 ng/ul) | NotI-HF | Cut Smart Buffer | ddH₂O | Total | |
17 | 6.0 ul | 0.5 ul | 2.0 ul | 11.5 ul | 20.0 ul |
18 | 13.0 ul | 0.5 ul | 2.0 ul | 4.5 ul | 20.0 ul |
37˚C 3h
65˚C 20’’ Heat Inactivation
Result: negative
GEL GÖRÜNTÜSÜ
Digestion of “pColA” (27.08.2015)
Digestion | |||||
---|---|---|---|---|---|
pColA (300 ng/ul) | NotI | Buffer | ddH₂O | Total | |
1x | 11.76 ul | 0.5 ul | 2.0 ul | 5.74 ul | 20.0 ul |
37˚C 3h
1.2 ul rSAP
37˚C 1h
20’’ Heat Inactivation
Colony PCR of “pCAGop-DsRed #12-2. Colony streak” (28.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
12x | 30.0 ul | 30.0 ul | 12.0 ul | 12.0 ul | 6.0 ul | 2.4 ul | 147.6 ul | 240.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: Negative
GEL GÖRÜNTÜSÜ
Colony PCR of „pSB1C3-T7 #5“ (28.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | VF | VR | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
16x | 40.0 ul | 40.0 ul | 16.0 ul | 16.0 ul | 13.0 ul | 3.2 ul | 196.8 ul | 325.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: #5-31 Bands were at the expected section
GEL GÖRÜNTÜSÜ
Cut Check of „pSB1C3-T7 #5-31“
Cut Check | ||||||
---|---|---|---|---|---|---|
DNA | EcoRI (FD) | PstI (FD) | Fast Digest Buffer | ddH₂O | Total | |
1x | ??????? | 1.0 ul | 1.0 ul | 2.0 ul | ????? | 20.0 ul |
GEL GÖRÜNTÜSÜ
Digestion of “pSB1C3 #9” (28.08.2015)
Digestion | ||||||
---|---|---|---|---|---|---|
DNA (500 ng/ul) | NotI | Cut Smart Buffer | ddH₂O | Total | ||
1x | ??????? | 1.0 ul | 2.0 ul | ????? | 20.0 ul |
3h was digested
Result: #9-5 Bands were at the expected section
GEL GÖRÜNTÜSÜ
Ligation of “pColA #9”
Ligation | ||||||
---|---|---|---|---|---|---|
pColA | #9 | T4 DNA Ligase | T4 DNA Ligase Buffer | ddH₂O | Total | |
1x | 7.5 ul | 9.2 ul | 1.0 ul | 2.0 ul | 0.3 ul | 20.0 ul |
2h was ligasted
Mini Prep of “pCAG-DsRed #12” (28.08.2015)
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/28/2015 7:23:31 PM | 0.5 | ng/ul | 0.010 | -0.006 | -1.58 | -1.46 | DNA | 50.00 |
2 | pcag op-dsred | biospec | 8/28/2015 7:24:38 PM | 92.3 | ng/ul | 1.846 | 1.105 | 1.67 | 0.79 | DNA | 50.00 |
3 | pcag op-dsred | biospec | 8/28/2015 7:26:20 PM | 67.4 | ng/ul | 1.348 | 0.702 | 1.92 | 1.35 | DNA | 50.00 |
Cut Check “pCAGop-DsRed #12”
Cut Check | ||||||
---|---|---|---|---|---|---|
DNA | EcoRI-HF | XhoI (NEB) | Cut Smart Buffer | ddH₂O | Total | |
1x | 3.7 ul | 0.5 ul | 0.5 ul | 2.0 ul | 13.3 ul | 20.0 ul |
Result: negative
GEL GÖRÜNTÜSÜ
Cut Check “pCAGop-DsRed #12” - Repeat
Cut Check | ||||||
---|---|---|---|---|---|---|
DNA | EcoRI-HF | XhoI (NEB) | Cut Smart Buffer | ddH₂O | Total | |
1x | 3.7 ul | 0.5 ul | 0.5 ul | 2.0 ul | 13.3 ul | 20.0 ul |
Result: negative
Colony PCR of „pTEToff and pTEToff N term“ (28.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | CMV rv | pTRE Luc fw | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
26x | 65.0 ul | 65.0 ul | 26.0 ul | 26.0 ul | 13.0 ul | 5.2 ul | 319.8 ul | 520.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: positive
GEL GÖRÜNTÜSÜ
Mini Prep of “pSB1C3-T7 TEV Protease” (28.09.2015)
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/28/2015 5:55:09 PM | 0.0 | ng/ul | -0.001 | -0.010 | 0.10 | -1.29 | DNA | 50.00 |
2 | tev psb1c3-t7/17 | biospec | 8/28/2015 5:56:45 PM | 81.8 | ng/ul | 1.637 | 0.848 | 1.93 | 2.08 | DNA | 50.00 |
3 | tev psb1c3-t7/18 | biospec | 8/28/2015 5:59:01 PM | 38.5 | ng/ul | 0.771 | 0.380 | 2.03 | 2.03 | DNA | 50.00 |
Mini Prep of “pSB1C3-T7 TEV Protease” (29.09.2015)
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/29/2015 8:08:11 PM | -0.8 | ng/ul | -0.016 | -0.019 | 0.83 | 0.68 | DNA | 50.00 |
2 | psb1c3-t7/tev17 | biospec | 8/29/2015 8:09:26 PM | 135.4 | ng/ul | 2.709 | 1.355 | 2.00 | 1.68 | DNA | 50.00 |
3 | psb1c3-t7/tev18 | biospec | 8/29/2015 8:11:06 PM | 229.1 | ng/ul | 4.581 | 2.193 | 2.09 | 2.40 | DNA | 50.00 |
Mini Prep of “pCAGop-DsRed #1-1” (30.08.2015)
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/30/2015 11:05:33 AM | 0.2 | ng/ul | 0.003 | -0.012 | -0.25 | -0.27 | DNA | 50.00 |
2 | pcag op-dsred | biospec | 8/30/2015 11:07:31 AM | 79.8 | ng/ul | 1.595 | 0.823 | 1.94 | 1.72 | DNA | 50.00 |
Cut Check of “pCAGop-DsRed #1-1”
Cut Check | ||||||
---|---|---|---|---|---|---|
pCAGop-DsRed (250 ng/ul) | EcoRI-HF | XhoI (NEB) | Cut Smart Buffer | ddH₂O | Total | |
1x | 3.2 ul | 0.5 ul | 0.5 ul | 2.0 ul | 13.8 ul | 20.0 ul |
37˚C 3h
Result: negative
GEL GÖRÜNTÜSÜ
Digestion of “pSB1C3-T7 TEV Protease” (30.08.2015)
Digestion | |||||
---|---|---|---|---|---|
Insert | NotI | Cut Smart Buffer | ddH₂O | Total | |
17 | 3.7 ul | 0.25 ul | 2.0 ul | 14.05 ul | 20.0 ul |
18 | 2.2 ul | 0.25 ul | 2.0 ul | 15.55 ul | 20.0 ul |
37˚C 3h
Result: Bands were at the expected section. Gel extraction was made.
GEL GÖRÜNTÜSÜ
Ligation of “pSB1C3-T7 TEV Protease”
Ligation | ||||||
---|---|---|---|---|---|---|
Vector (60 ng/ul) | Insert (29.11 ng/ul) | T4 DNA Buffer | T4 DNA Buffer Ligase | ddH₂O | Total | |
1x | 7.0 ul | 8.7 ul | 2.0 ul | 1.0 ul | 1.3 ul | 20.0 ul |
22˚C 1.5h
Transformation was made.
Gel Extraction
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 8/30/2015 5:28:51 PM | 0.0 | ng/ul | -0.001 | -0.005 | 0.15 | 0.03 | DNA | 50.00 |
2 | psb1c3-t7 | biospec | 8/30/2015 5:28:05 PM | 5.6 | ng/ul | 0.113 | 0.061 | 1.85 | 0.17 | DNA | 50.00 |
Colony PCR of “pColA – TEV” (31.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pColA fw | pColA rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
16x | 40.0 ul | 40.0 ul | 16.0 ul | 16.0 ul | 8.0 ul | 3.2 ul | 196.8 ul | 320.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
Colony PCR of “pColA – TEV” – Repeat (31.08.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pColA fw | pColA rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
22x | 55.0 ul | 55.0 ul | 22.0 ul | 22.0 ul | 11.0 ul | 3.2 ul | 270.6 ul | 440.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
Digestion of “PsicA for pSB1C3 and invF-sicA for ColA” (31.08.2015)
Digestion of “PsicA and invF” | |||||||||
---|---|---|---|---|---|---|---|---|---|
DNA | EcoRI-HF | PstI | XbaI | SpeI | 2.1 Buffer | Cut Smart Buffer | ddH₂O | Total | |
invF-sicA | 10.0 ul | - | - | 1.0 ul | 1.0 ul | - | 2.0 ul | 6.0 ul | 20.0 ul |
PsicA | 10.0 ul | 1.0 ul | 1.0 ul | - | - | 2.0 ul | - | 6.0 ul | 20.0 ul |
37˚C 10h
82˚C 20’ Heat Inactivation
Digestion of “pSB1C3-RFP and pColA-NotI” | |||||||||
---|---|---|---|---|---|---|---|---|---|
DNA | EcoRI (FD) | PstI (FD) | XbaI (FD) | SpeI (FD) | Fast Digest Buffer | ddH₂O | Total | ||
pSB1C3-RFP (1000 ng/ul) | 2.5 ul | 1.0 ul | 1.0 ul | - | - | 2.0 ul | 13.5 ul | 20.0 ul | |
pColA-NotI (500 ng/ul) | 5.0 ul | - | - | 1.0 ul | 1.0 ul | 2.0 ul | 11.0 ul | 20.0 ul |
September
02.09.2015 Wiki meetings continue, we are making decisions together. 06.09.2015 What will we be wearing at the Giant Jamboree? Thinking about how to design our T-shirts. 08.09.2015 Team pictures mustn’t be left ends. 11.09.2015 We taught our little friends synthetic biology. Those kids were really funny when they came first, they were covering their faces because they were scared of being contaminated. 13.09.2015 Coloring our drawings is complete. They are better than original now. We still have wiki meetings, there are only 5 days to wiki-freeze!
Ligation of “ColA and pSB1C3” (01.09.2015)
Ligation | |||||||
---|---|---|---|---|---|---|---|
Vector (100 ng/ul) | invF-sicA | psicA | T4 DNA Ligase | T4 DNA Ligase Buffer | ddH₂O | Total | |
ColA | 8.5 ul | 17.5 ul | - | 1.0 ul | 2.0 ul | - | 30.0 ul |
pSB1C3 | 9.44 ul | - | 13.55 ul | 1.0 ul | 2.0 ul | 3.0 ul | 30.0 ul |
22˚C 2h
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/1/2015 4:10:17 AM | 0.3 | ng/ul | 0.006 | -0.007 | -0.88 | -0.78 | DNA | 50.00 |
2 | colA ext | biospec | 9/1/2015 4:11:28 AM | 11.7 | ng/ul | 0.235 | 0.132 | 1.78 | 0.08 | DNA | 50.00 |
3 | cola clean-up | biospec | 9/1/2015 4:12:56 AM | 7.2 | ng/ul | 0.143 | 0.078 | 1.84 | 0.06 | DNA | 50.00 |
4 | psb1c3 ext | biospec | 9/1/2015 4:14:40 AM | 10.6 | ng/ul | 0.213 | 0.113 | 1.88 | 0.12 | DNA | 50.00 |
Transformation will be made.
Ligation of “pCAGop-DsRed” (01.09.2015)
Ligation | |||||||
---|---|---|---|---|---|---|---|
pCAG | pTRE #12 | T4 DNA Ligase | T4 DNA Ligase Buffer | ddH₂O | Total | ||
1x | 0.3 ul | 4.0 ul | 1.0 ul | 1.0 ul | 13.7 ul | 20.0 ul |
Transformation with BL21.
Digestion of “pSB1C3-T7 TEV and pColA” (01.09.2015)
Digestion | |||||||
---|---|---|---|---|---|---|---|
DNA | XhoI-HF | SpeI-HF | Cut Smart Buffer | RNAse | ddH₂O | Total | |
17 (Hand Made Solution) | 10.0 ul | 0.25 ul | 0.25 ul | 2.0 ul | 0.2 ul | 7.3 ul | 20.0 ul |
18 (Hand Made Solution) | 10.0 ul | 0.25 ul | 0.25 ul | 2.0 ul | 0.2 ul | 7.3 ul | 20.0 ul |
17 (Midi Prep Solution) | 10.0 ul | 0.25 ul | 0.25 ul | 2.0 ul | - | 7.5 ul | 20.0 ul |
18 (Midi Prep Solution) | 10.0 ul | 0.25 ul | 0.25 ul | 2.0 ul | - | 7.5 ul | 20.0 ul |
pColA (1000 ng/ul) | 9.0 ul | 0.5 ul | 0.5 ul | 2.0 ul | - | 8.0 ul |
37˚C 3h
80˚C 20’ Heat Inactivation
Cut Check was made.
Görüntü
Result: pColA Bands were at the expected section. Midi Prep Solution is better.
Colony PCR of “pCAGop-DsRed and pCMV-lacO” (02.09.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
16x | 40.0 ul | 40.0 ul | 16.0 ul | 16.0 ul | 8.0 ul | 3.2 ul | 196.8 ul | 320.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative. It will be repeated.
GEL GÖRÜNTÜSÜ
Colony PCR of “invF-sicA and PsicA” (02.09.2015)
Result: PsicA: 1, 2, 6, 8, 9, 10, 11 Liquid Culture will be made.
GEL GÖRÜNTÜSÜ
Digestion of “invF-sicA for pSB1C3 and ColA” (02.09.2015)
DNA 10.0 ul + rSAP 1.0 ul / 37˚C 1h / 80˚C 20’
Digestion | |||||||||
---|---|---|---|---|---|---|---|---|---|
DNA | EcoRI-HF | PstI | XbaI | SpeI | 2.1 Buffer | Cut Smart Buffer | ddH₂O | Total | |
pSB1C3 | 10.0 ul | 0.4 ul | 0.4 ul | - | - | 2.0 ul | - | 7.2 ul | 20.0 ul |
ColA | 10.0 ul | - | - | 0.4 ul | 0.4 ul | - | 2.0 ul | 7.2 ul | 20.0 ul |
37˚C overnight
Mini Prep of “PsicA – pSB1C3” (03.08.2015)
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/3/2015 8:34:29 PM | 0.8 | ng/ul | 0.016 | 0.000 | 116.17 | 0.85 | DNA | 50.00 |
2 | psb1c3-psica 1 | biospec | 9/3/2015 8:35:32 PM | 130.5 | ng/ul | 2.610 | 1.370 | 1.91 | 2.04 | DNA | 50.00 |
3 | psb1c3-psica 2 | biospec | 9/3/2015 8:36:34 PM | 140.9 | ng/ul | 2.818 | 1.457 | 1.93 | 2.16 | DNA | 50.00 |
4 | psb1c3-psica 6 | biospec | 9/3/2015 8:37:28 PM | 178.5 | ng/ul | 3.569 | 1.864 | 1.92 | 2.20 | DNA | 50.00 |
5 | psb1c3-psica 8 | biospec | 9/3/2015 8:38:24 PM | 98.7 | ng/ul | 1.974 | 1.021 | 1.93 | 2.13 | DNA | 50.00 |
6 | psb1c3-psica 9 | biospec | 9/3/2015 8:39:21 PM | 237.0 | ng/ul | 4.741 | 2.476 | 1.91 | 2.13 | DNA | 50.00 |
7 | psb1c3-psica 10 | biospec | 9/3/2015 8:40:35 PM | 109.0 | ng/ul | 2.181 | 1.127 | 1.94 | 2.05 | DNA | 50.00 |
8 | biospec | 9/3/2015 8:41:51 PM | 109.5 | ng/ul | 2.189 | 1.134 | 1.93 | 1.97 | DNA | 50.00 | |
9 | blank | biospec | 9/3/2015 8:43:10 PM | 1.3 | ng/ul | 0.025 | 0.003 | 9.26 | 0.73 | DNA | 50.00 |
10 | psb1c3-psica 11 | biospec | 9/3/2015 8:44:14 PM | 50.4 | ng/ul | 1.008 | 0.518 | 1.95 | 1.97 | DNA | 50.00 |
Cut Check of “PsicA – pSB1C3”
Cut Check | ||||||
---|---|---|---|---|---|---|
DNA | EcoRI (FD) | PstI (FD) | Fast Digest Buffer | ddH₂O | Total | |
1 | 1.9 ul | 0.35 ul | 0.35 ul | 2.0 ul | 15.4 ul | 20.0 ul |
2 | 1.8 ul | 0.35 ul | 0.35 ul | 2.0 ul | 15.5 ul | 20.0 ul |
6 | 1.4 ul | 0.35 ul | 0.35 ul | 2.0 ul | 15.9 ul | 20.0 ul |
8 | 2.5 ul | 0.35 ul | 0.35 ul | 2.0 ul | 14.8 ul | 20.0 ul |
9 | 1.1 ul | 0.35 ul | 0.35 ul | 2.0 ul | 16.2 ul | 20.0 ul |
10 | 2.3 ul | 0.35 ul | 0.35 ul | 2.0 ul | 15.0 ul | 20.0 ul |
11 | 5.0 ul | 0.35 ul | 0.35 ul | 2.0 ul | 12.3 ul | 20.0 ul |
37˚C 35’
Ligation of “invF and pSB1C3/pColA” (03.08.2015)
Ligation | |||||||
---|---|---|---|---|---|---|---|
Vector (100 ng/ul) | invF/sicA | invF/sicA | T4 DNA Ligase | T4 DNA Ligase Buffer | ddH₂O | Total | |
ColA (-) | 11.6 ul | - | - | 1.0 ul | 3.2 ul | 16.6 ul | 32.2 ul |
ColA (+) | 11.6 ul | 14.40 ul | - | 3.2 ul | 3.4 ul | 2.2 ul | 34.8 ul |
pSB1C3 | 9.4 ul | - | 14.40 ul | 1.0 ul | 3.0 ul | 2.2 ul | 30.0 ul |
RT 1h
Transformation will be made.
Colony PCR of “pCMV-Lacop-DsRed and pCAGop-DsRed” (04.09.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
15x | 37.5 ul | 37.5 ul | 15.0 ul | 15.0 ul | 7.5 ul | 3.0 ul | 184.5 ul | 300.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: Bands were at the expected section. Liquid Culture was made.
GEL GÖRÜNTÜSÜ
Mini Prep of „pCMV-Lacop-DsRed and pCAGop-DsRed“ (05.09.2015)
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/5/2015 10:42:13 AM | -0.1 | ng/ul | -0.003 | -0.015 | 0.18 | 0.15 | DNA | 50.00 |
2 | pcmv-lacop-dsred 1 | biospec | 9/5/2015 10:44:11 AM | 68.4 | ng/ul | 1.369 | 0.690 | 1.98 | 1.76 | DNA | 50.00 |
3 | pcmv-lacop-dsred 2 | biospec | 9/5/2015 10:45:12 AM | 51.2 | ng/ul | 1.024 | 0.499 | 2.05 | 1.77 | DNA | 50.00 |
4 | pcmv-lacop-dsred 3 | biospec | 9/5/2015 10:46:09 AM | 78.5 | ng/ul | 1.570 | 0.793 | 1.98 | 1.98 | DNA | 50.00 |
5 | pcmv-lacop-dsred 4 | biospec | 9/5/2015 10:47:04 AM | 47.7 | ng/ul | 0.954 | 0.460 | 2.07 | 1.95 | DNA | 50.00 |
6 | Pcagop-dsred 1 | biospec | 9/5/2015 10:48:08 AM | 499.6 | ng/ul | 9.991 | 5.290 | 1.89 | 2.20 | DNA | 50.00 |
Cut Check of „pCMV-Lacop-DsRed and pCAGop-DsRed“
Cut Check | |||||||
---|---|---|---|---|---|---|---|
DNA | BamHI | XhoI | EcoRI | Cut Smart Buffer | ddH₂O | Total | |
1 | 4.4 ul | 0.5 ul | 0.5 ul | - | 2.0 ul | 12.6 ul | 20.0 ul |
2 | 5.9 ul | 0.5 ul | 0.5 ul | - | 2.0 ul | 11.1 ul | 20.0 ul |
3 | 3.9 ul | 0.5 ul | 0.5 ul | - | 2.0 ul | 13.1 ul | 20.0 ul |
4 | 6.3 ul | 0.5 ul | 0.5 ul | - | 2.0 ul | 10.7 ul | 20.0 ul |
pCAGop-DsRed | 1.0 ul | 0.5 ul | - | 0.5 ul | 2.0 ul | ||
16.0 ul | 20.0 ul |
37˚C 3h
Result: 1-2-3-4 is positive.
GEL GÖRÜNTÜSÜ
Cut Check of “invF and sicA” (07.09.2015)
Cut Check | |||||||
---|---|---|---|---|---|---|---|
DNA (250 ng/ul) | EcoRI (FD) | PstI (FD) | Fast Digest Buffer | ddH₂O | Total | ||
1x | 5.0 ul | 0.5 ul | 0.5 ul | 2.0 ul | 12.0 ul | 20.0 ul |
37˚C 35’
GEL GÖRÜNTÜSÜ
Mini Prep
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/7/2015 6:34:56 PM | -0.1 | ng/ul | -0.003 | -0.005 | 0.60 | 0.11 | DNA | 50.00 |
2 | #18-11 | biospec | 9/7/2015 6:36:20 PM | 79.8 | ng/ul | 1.597 | 0.843 | 1.89 | 1.93 | DNA | 50.00 |
3 | #18-2 | biospec | 9/7/2015 6:37:48 PM | 74.9 | ng/ul | 1.497 | 0.787 | 1.90 | 2.01 | DNA | 50.00 |
4 | elution blank | biospec | 9/7/2015 6:39:44 PM | 0.3 | ng/ul | 0.005 | -0.004 | -1.51 | 0.68 | DNA | 50.00 |
5 | #18-5 elution | biospec | 9/7/2015 6:40:49 PM | 57.7 | ng/ul | 1.154 | 0.604 | 1.91 | 1.91 | DNA | 50.00 |
Digestion of “#18-2,5”
Digestion | ||||||
---|---|---|---|---|---|---|
DNA (500 ng/ul) | XbaI | SpeI-HF | Cut Smart Buffer | ddH₂O | Total | |
#18-2 | 6.66 ul | 0.4 ul | 0.4 ul | 2.0 ul | 8.54 ul | 20.0 ul |
#18-5 | 8.66 ul | 0.4 ul | 0.4 ul | 2.0 ul | 10.54 ul | 20.0 u |
37˚C 40’
Ligation
Ligation | ||||||
---|---|---|---|---|---|---|
pSB1C3-invF (175 ng/ul) | ColA (100 ng/ul) | T4 DNA Ligase | T4 DNA Ligase Buffer | ddH₂O | Total | |
1 | 7.0 ul | - | 1.0 ul | 2.0 ul | 9.17 ul | 20.0 ul |
2 | - | 7.0 ul | 1.0 ul | 2.0 ul | 5.0 ul | 20.0 ul |
Transformation will be made.
Mini Prep of “pSB1C3 – TEV”
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/8/2015 10:39:08 AM | -0.1 | ng/ul | -0.003 | -0.009 | 0.30 | 0.13 | DNA | 50.00 |
2 | psb1c3-tev 17 | biospec | 9/8/2015 10:40:15 AM | 125.5 | ng/ul | 2.510 | 1.331 | 1.89 | 2.16 | DNA | 50.00 |
3 | psb1c3-tev 18 | biospec | 9/8/2015 10:42:45 AM | 111.9 | ng/ul | 2.238 | 1.195 | 1.87 | 2.14 | DNA | 50.00 |
Digestion of “pSB1C3 – TEV 17/18 and pColA-Toehold”
Digestion of “pSB1C3 – TEV 17/18” | ||||||
---|---|---|---|---|---|---|
DNA (1000 ng/ul) | XbaI (FD) | SpeI (FD) | Fast Digest Buffer | ddH₂O | Total | |
17 | 8.0 ul | 1.0 ul | 1.0 ul | 2.0 ul | 8.0 ul | 20.0 ul |
18 | 9.0 ul | 1.0 ul | 1.0 ul | 2.0 ul | 7.0 ul | 20.0 ul |
37˚C 1h
Digestion of “pColA-Toehold” | ||||||
---|---|---|---|---|---|---|
DNA (1000 ng/ul) | XbeI (FD) | SpeI (FD) | Fast Digest Buffer | ddH₂O | Total | |
1x | 16.0 ul | 1.0 ul | 1.0 ul | 2.0 ul | - | 20.0 ul |
37˚C 1h
Colony PCR of “ pSB1C3-TEV 17/18” (08.09.2015)
Colony PCR | |||||||||
---|---|---|---|---|---|---|---|---|---|
MgCl₂ | (NH₄)2SO₄ | pTRE Luc fw | SV40 rv | dNTP | Tag | ddH₂O | DNA | Total | |
1x | 2.5 ul | 2.5 ul | 1.0 ul | 1.0 ul | 0.5 ul | 0.2 ul | 12.3 ul | 5.0 ul | 25.0 ul |
25x | 61.5 ul | 61.5 ul | 25.0 ul | 25.0 ul | 12.5 ul | 5.0 ul | 307.5 ul | 498.0 ul |
Cycling | ||||||
---|---|---|---|---|---|---|
95˚C | 95˚C | 56˚C | 72˚C | 72˚C | Cycle | |
Time | 5’ | 30’’ | 30’’ | 1.5’ | 5’ | 35x |
Result: negative
GEL GÖRÜNTÜSÜ
Digestion of “pColA / pSB1C3 #9 / pColA-Toehold #2-8 / pSB1C3 T7 #10-17 / #18” (10.11.2015)
Digestion of | ||||||
---|---|---|---|---|---|---|
DNA (1000 ng/ul) | XbaI (FD) | SpeI (FD) | Cut Smart Buffer | ddH₂O | Total | |
pColA | 4.5 ul | 0.5 ul | 0.5 ul | 2.0 ul | 12.5 ul | 20.0 ul |
pSB1C3 #9 | 1.7 ul | 0.5 ul | 0.5 ul | 2.0 ul | 15.3 ul | 20.0 ul |
pColA-Toehold #2-8 | 16.0 ul | 0.5 ul | 0.5 ul | 2.0 ul | 1.0 ul | 20.0 ul |
pSB1C3 T7 #10-17 | 3.3 ul | 0.5 ul | 0.5 ul | 2.0 ul | 13.7 ul | 20.0 ul |
#18 | 3.0 ul | 0.5 ul | 0.5 ul | 2.0 ul | 14.0 ul | 20.0 ul |
37˚C 3h<7p> <p>pColA-Toehold #2-8: 1ul rSAP / 37˚C 1h / 80˚C 20’
Result: pColA and #10 gel extraction will be made.
GEL GÖRÜNTÜLERI
Gel extraction
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/10/2015 7:29:27 PM | -0.2 | ng/ul | -0.003 | -0.009 | 0.37 | 0.28 | DNA | 50.00 |
2 | psb1c3-#9 | biospec | 9/10/2015 7:31:01 PM | 308.0 | ng/ul | 6.160 | 3.182 | 1.94 | 2.10 | DNA | 50.00 |
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/10/2015 8:13:13 PM | 0.3 | ng/ul | 0.005 | -0.003 | -1.70 | -2.25 | DNA | 50.00 |
2 | psb1c3 t7-10 (17) | biospec | 9/10/2015 8:14:54 PM | 152.2 | ng/ul | 3.045 | 1.612 | 1.89 | 2.25 | DNA | 50.00 |
3 | psb1c3 t7-10 (18) | biospec | 9/10/2015 8:16:02 PM | 164.6 | ng/ul | 3.291 | 1.737 | 1.90 | 2.23 | DNA | 50.00 |
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/11/2015 10:05:05 PM | 0.1 | ng/ul | 0.002 | -0.017 | -0.13 | -0.11 | DNA | 50.00 |
2 | pcola (notI) | biospec | 9/11/2015 10:07:08 PM | 40.4 | ng/ul | 0.807 | 0.399 | 2.03 | 1.75 | DNA | 50.00 |
3 | psb1c3 #9 | biospec | 9/11/2015 10:08:33 PM | 76.0 | ng/ul | 1.520 | 0.789 | 1.93 | 2.17 | DNA | 50.00 |
4 | psb1c3-t7 #10 | biospec | 9/11/2015 10:09:55 PM | 71.5 | ng/ul | 1.431 | 0.721 | 1.99 | 2.15 | DNA | 50.00 |
5 | blank | biospec | 9/11/2015 10:14:04 PM | -0.4 | ng/ul | -0.007 | -0.020 | 0.35 | 0.32 | DNA | 50.00 |
6 | psb1c3-t7 #10 | biospec | 9/11/2015 10:15:01 PM | 70.4 | ng/ul | 1.408 | 0.727 | 1.94 | 2.16 | DNA | 50.00 |
# | Sample ID | User name | Date and Time | Nucleic Acid Conc. | Unit | A260 | A280 | 260/280 | 260/230 | Sample Type | Factor |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | blank | biospec | 9/11/2015 2:24:30 AM | 0.6 | ng/ul | 0.011 | 0.002 | 4.95 | 1.18 | DNA | 50.00 |
2 | Pcola-2 | biospec | 9/11/2015 2:26:25 AM | 145.0 | ng/ul | 2.901 | 1.398 | 2.08 | 2.04 | DNA | 50.00 |