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''By Léa & Manhaz''
+ We have decided to use the plasmid coding dCas9 Nm to amplify the fragement corresponding g-block 2.2.
200 mL of transformed DH5a cells with Adgene dCas9 Nm plasmids (clone 1 and 2) cultures were extracted usind a Midi Prep kit. It was resuspended into 50µL of sterile water.
200 mL of transformed DH5a cells with Adgene dCas9 Nm plasmids (clone 1 and 2) cultures were extracted usind a Midi Prep kit. It was resuspended into 50µL of sterile water.
+
+ The plasmid then reserved in -20.
====PCR Q5 on plasmid ====
====PCR Q5 on plasmid ====
Revision as of 12:52, 20 September 2016
Wednesday 24th August Lab work Visualization Gel Electrophoresis of Gibson Assembly's products(fragment 3-4) and GFP 1-9 cloned in PSB1C3 By Terrence
Colony PCR of 12 colonies transformed with Gibson Assembly products (fragment 3-4).
Colony PCR of 12 colonies containing the GFP 1-9.
- Clones 2, 4, 7 and 11 are as expected.
pPS16_002 (1.2) clones 8 and 14 and pPS16_014(SgRna Nm) clone 12 Extraction By Terrence
The extraction was carried out following the usual protocol .
Nano drop :
concentration (ng/µL)
260/230
260/280
1.2.14
47.81
1.64
1.84
1.2.8
834.5
2.40
1.99
SgRna Nm 12
124.21
2.00
1.89
Transformation of gblockFRB in pJET and gblock Sg_RNA_NM in pJET By Terrence
Component
Volume (µL)
Reaction Buffer
10
PCR Product (FRB or Nm)
1
pJET1.2/blunt
1
water, nuclease free
7
T4 DNA Ligase
1
Then we transformed 2µL of ligation product into 50 µL of DH5a competent cells, following the usual protocol .
Transformation products were plated with three different volume : 50µL , 150µl and 350µL. 350µL only for the control.
By Léa & Manhaz
We have decided to use the plasmid coding dCas9 Nm to amplify the fragement corresponding g-block 2.2.
200 mL of transformed DH5a cells with Adgene dCas9 Nm plasmids (clone 1 and 2) cultures were extracted usind a Midi Prep kit. It was resuspended into 50µL of sterile water.
The plasmid then reserved in -20.
PCR Q5 on plasmid By Léa & Manhaz
