Difference between revisions of "Team:UCLA/Notebook/Spider Silk Genetics/12 August 2015"

(→‎BsaI Digestion of M1AB, BC and CA)
 
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| 4 uL
 
| 4 uL
 
|}
 
|}
 +
 +
{| class="wikitable"
 +
| 95C
 +
| 3 min
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|-
 +
| 95C
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| 30 sec
 +
|-
 +
| 72C
 +
| 25 sec (M1SeqAB)/ 1min 40 sec (M1-9)
 +
|-
 +
|
 +
| x30
 +
|-
 +
| 72C
 +
| 5 min
 +
|-
 +
| 12C
 +
| Hold
 +
|}
 +
 +
[[File:ColonyPCR0812.jpg |800px|thumb|left|Lane 3-6: M1-9(T7) Colony 1-4. Lane 7-10: M1SeqAB1 Colony 1-4. Lane 11-14: M1SeqAB2 Colony 1-4]]
 +
 +
Grew liquid culture for
 +
* all 4 colonies of M1-9 (T7); they all showed band close to the expected size ~1200-1300bp but they are slightly different so decided to sequence all of them.
 +
* colonies 1-3 for M1SeqAB1
 +
* none for M1SeqAB2 since we probably do not need this seq construct
 +
 +
== BsaI Digestion of M1AB, BC and CA==
 +
 +
{| class="wikitable"
 +
|
 +
| 1AB (553.88 ng/uL)
 +
| 1BC (1477.18 ng/uL)
 +
| 1CA (648 ng/ uL)
 +
|-
 +
|Custmart
 +
| 5 uL
 +
| 5 uL
 +
| 5 uL
 +
|-
 +
| DNA
 +
| 7.2 uL
 +
| 2.7 uL
 +
| 6.2 uL
 +
|-
 +
| ddH2O
 +
| 33.8 uL
 +
| 38.3 uL
 +
| 34.8 uL
 +
|-
 +
| BsaI
 +
| 4 uL
 +
| 4 uL
 +
| 4 uL
 +
|}
 +
 +
50C for 2 hr
 +
65C for 20 min
 +
 +
[[File:BsaIDig0812.jpg|500px]]
 +
 +
Gel purified samples

Latest revision as of 06:45, 13 August 2015

iGEM UCLA




08/12/2015

Colony PCR for M1 SwqAB 1 & 2 and M1-9(T7)

2X Taq MM 6.25 uL
VF Primer 0.625 uL
VR Primer 0.625 uL
Template 1 uL
ddH2O 4 uL
95C 3 min
95C 30 sec
72C 25 sec (M1SeqAB)/ 1min 40 sec (M1-9)
x30
72C 5 min
12C Hold
Lane 3-6: M1-9(T7) Colony 1-4. Lane 7-10: M1SeqAB1 Colony 1-4. Lane 11-14: M1SeqAB2 Colony 1-4

Grew liquid culture for

  • all 4 colonies of M1-9 (T7); they all showed band close to the expected size ~1200-1300bp but they are slightly different so decided to sequence all of them.
  • colonies 1-3 for M1SeqAB1
  • none for M1SeqAB2 since we probably do not need this seq construct

BsaI Digestion of M1AB, BC and CA

1AB (553.88 ng/uL) 1BC (1477.18 ng/uL) 1CA (648 ng/ uL)
Custmart 5 uL 5 uL 5 uL
DNA 7.2 uL 2.7 uL 6.2 uL
ddH2O 33.8 uL 38.3 uL 34.8 uL
BsaI 4 uL 4 uL 4 uL

50C for 2 hr 65C for 20 min

BsaIDig0812.jpg

Gel purified samples