Difference between revisions of "Team:Paris Saclay/Notebook/August/24"

(→‎Migration gel of Gibson (fragment 3 + fragment 4) and GFP in PSB1C3)
(→‎Transformation of pPS16_010 (FRB) in pJET and pPS16_014 (SgRnaNm) in pJET)
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Then we transformed 2µL of ligation product into 50 µL of competent cells, following the [[Team:Paris_Saclay/Experiments#Heat_shock_competent_cells|usual protocol]].
 
Then we transformed 2µL of ligation product into 50 µL of competent cells, following the [[Team:Paris_Saclay/Experiments#Heat_shock_competent_cells|usual protocol]].
  
 +
Transformation products were plated in duplicate : 50µL , 150µl and 350µL. 350µL only for the control.
  
 
{{Team:Paris_Saclay/notebook_footer}}
 
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 16:22, 24 August 2016

Wednesday 24th August

Lab work

Visualization

Gel Electrophoresis of Gibson (fragment 3 + fragment 4) and GFP in PSB1C3

By Terrence


Clone 6 and 8 seems correct for the Gibson.


Clone 2-4-7-11 are correct.

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.


Result of the extraction


Nano drop :

size (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 pPS16_010 (FRB) in pJET and pPS16_014 (SgRnaNm) in pJET

By Terrence


First we made a ligation with

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 competent cells, following the usual protocol.

Transformation products were plated in duplicate : 50µL , 150µl and 350µL. 350µL only for the control.





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