Difference between revisions of "Team:Paris Saclay/Notebook/August/20"
(Created page with "=Thursday 20th August= ==Lab Work== ===Digestion for check=== ''by Audrey'' Biobricks: * BBa_K115017 (control +) * BBa_J23101 + GFP (control -) * BBa_K1707000 #2, #3 and #4 * ...") |
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| + | {{Team:Paris_Saclay/notebook_header}} | ||
=Thursday 20th August= | =Thursday 20th August= | ||
==Lab Work== | ==Lab Work== | ||
| − | |||
===Digestion for check=== | ===Digestion for check=== | ||
''by Audrey'' | ''by Audrey'' | ||
| − | |||
* BBa_K115017 (control +) | * BBa_K115017 (control +) | ||
* BBa_J23101 + GFP (control -) | * BBa_J23101 + GFP (control -) | ||
| Line 25: | Line 24: | ||
** 2 µL Buffer 3 10x (Biolabs) | ** 2 µL Buffer 3 10x (Biolabs) | ||
** 5 µL H2O | ** 5 µL H2O | ||
| − | |||
===Electrophoresis to check PCR products=== | ===Electrophoresis to check PCR products=== | ||
| Line 36: | Line 34: | ||
We can conclude that all clones aren't OK | We can conclude that all clones aren't OK | ||
| − | |||
===Purification=== | ===Purification=== | ||
''by Pauline'' | ''by Pauline'' | ||
| − | |||
* BBa_K1707022 #1 | * BBa_K1707022 #1 | ||
* BBa_K1707023 #1 | * BBa_K1707023 #1 | ||
| Line 55: | Line 51: | ||
Digestion products: | Digestion products: | ||
| − | |||
* BBa_K115017 (control +) | * BBa_K115017 (control +) | ||
* BBa_J23101 + GFP (control -) | * BBa_J23101 + GFP (control -) | ||
| Line 69: | Line 64: | ||
Quantification: | Quantification: | ||
| − | |||
* BBa_K1707022 #1: ? | * BBa_K1707022 #1: ? | ||
* BBa_K1707023 #1: nothing | * BBa_K1707023 #1: nothing | ||
| Line 80: | Line 74: | ||
''by Pauline'' | ''by Pauline'' | ||
| − | + | * BBa_K1707004 #2 | |
In 5mL LB + 5µL Ampicillin | In 5mL LB + 5µL Ampicillin | ||
| − | |||
| − | |||
===Ligation=== | ===Ligation=== | ||
''by Pauline'' | ''by Pauline'' | ||
| − | BBa_K1707037= BBa_K1707028 + BBa_K1707034 | + | * BBa_K1707037 = BBa_K1707028 + BBa_K1707034 |
Mix: | Mix: | ||
| Line 99: | Line 91: | ||
Incubation 4°C, ON | Incubation 4°C, ON | ||
| − | |||
===Quantification=== | ===Quantification=== | ||
| Line 109: | Line 100: | ||
We can't see anything | We can't see anything | ||
| − | |||
| − | |||
| − | |||
'''Member present:''' | '''Member present:''' | ||
Revision as of 07:57, 15 September 2015
Contents
Thursday 20th August
Lab Work
Digestion for check
by Audrey
- BBa_K115017 (control +)
- BBa_J23101 + GFP (control -)
- BBa_K1707000 #2, #3 and #4
- BBa_K1707013 #1
- BBa_K1707030 #1
- BBa_K1707019 #1
- BBa_K1707020 #2
- BBa_K1707021 #2
- BBa_K1707027 #1
- BBa_K1707035 #1
- BBa_K1707036 #1
Mix for each tub:
- 2 µL plasmid
- 1 µL SpeI (Biolabs)
- 1 µL BspE1 (Biolabs)
- 2 µL Buffer 3 10x (Biolabs)
- 5 µL H2O
Electrophoresis to check PCR products
by Audrey
Agarose gel 1%, Migration 90V
Biobricks:
- BBa_K1707031 #7 to #26
We can conclude that all clones aren't OK
Purification
by Pauline
- BBa_K1707022 #1
- BBa_K1707023 #1
- BBa_K1707004 #2
- BBa_K1707034 #1
- BBa_K1707028 #1
With Macherey Nagel purification kit
Electrophoresis
by Pauline
Digestion products:
- BBa_K115017 (control +)
- BBa_J23101 + GFP (control -)
- BBa_K1707000 #2, #3 and #4
- BBa_K1707013 #1
- BBa_K1707030 #1
- BBa_K1707019 #1
- BBa_K1707020 #2
- BBa_K1707021 #2
- BBa_K1707027 #1
- BBa_K1707035 #1
- BBa_K1707036 #1
Quantification:
- BBa_K1707022 #1: ?
- BBa_K1707023 #1: nothing
- BBa_K1707004 #2: nothing
- BBa_K1707034 #1: 20 µg/µL
- BBa_K1707028 #1: 10 µg/µL
New culture
by Pauline
- BBa_K1707004 #2
In 5mL LB + 5µL Ampicillin
Ligation
by Pauline
- BBa_K1707037 = BBa_K1707028 + BBa_K1707034
Mix:
- 8 µL BBa_K1707028
- 10 µL BBa_K1707034
- 2 µL Ligase
- 2,5 µL Buffer
- 2,5 µL H2O
Incubation 4°C, ON
Quantification
by Pauline
BIobricks:
- BBa_K1707022 #1
- BBa_K1707023 #1
We can't see anything
Member present:
- Instructors: Claire
- Students: Pauline and Audrey
Contact
eigem.parissaclay AT gmail DOT com
t@iGEMParisSaclay
aUniversité Paris Sud
a91405 Orsay, France
Copyright © 2015 iGEM Paris Saclay. All rights reserved All inspirations encouraged.
*This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.
eigem.parissaclay AT gmail DOT com
t@iGEMParisSaclay
aUniversité Paris Sud
a91405 Orsay, France
Copyright © 2015 iGEM Paris Saclay. *This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.