Difference between revisions of "Team:Paris Saclay/Notebook/August/2"

(Culture of DH5alp|pPS16_003, DH5alp|pPS16_004, DH5alp|pPS16_006, DH5alp|pPS16_007 and DH5alp|pPS16_008)
(Lab work)
 
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{{Team:Paris_Saclay/notebook_header}}
 
{{Team:Paris_Saclay/notebook_header}}
= Tuesday 2<sup>st</sup> August=
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= Tuesday 2<sup>nd</sup> August=
==Lab work==
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===Visualization===
 
===Visualization===
 
==== DreamTaq PCR on DH5alp|pPS16_001, DH5alp|pPS16_002, DH5alp|pPS16_005, DH5alp|pPS16_009, DH5alp|pPS16_0010, DH5alp|pPS16_011, DH5alp|pPS16_012, DH5alp|pPS16_013, DH5alp|pPS16_014, DH5alp|pPS16_015 ====
 
==== DreamTaq PCR on DH5alp|pPS16_001, DH5alp|pPS16_002, DH5alp|pPS16_005, DH5alp|pPS16_009, DH5alp|pPS16_0010, DH5alp|pPS16_011, DH5alp|pPS16_012, DH5alp|pPS16_013, DH5alp|pPS16_014, DH5alp|pPS16_015 ====
 
''By Charlène, Mathilde, Laetitia, Caroline and Léa''
 
''By Charlène, Mathilde, Laetitia, Caroline and Léa''
  
A Low fidelity PCR was carried out following the usual [[Team:Paris_Saclay/Experiments#Polymerase_chain_reaction|protocol]] and using the universal puc19 [[Team:Paris_Saclay/Experiments#primers|primers]]. 6 clones for each transformations were tested.
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A Low fidelity PCR was carried out following the usual [[Team:Paris_Saclay/Experiments#taqPCR|protocol]] and using the puc19 [[Team:Paris_Saclay/Experiments#primers|primers]]. 6 clones for each transformations were tested.
  
Each clone was re-plated and put at 37°C.
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Each clone was spread and put at 37°C.
 
We also did a liquid culture for each clone in 3mL of LB and Ampicilin(37°C and 180 rpm).
 
We also did a liquid culture for each clone in 3mL of LB and Ampicilin(37°C and 180 rpm).
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[[File:T--Paris Saclay--160802 gel pcr 1.jpg|400px|thumb|right|Results]]
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[[File:T--Paris Saclay--160802 gel pcr 2.jpg|400px|thumb|right|Results]]
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[[File:T--Paris Saclay--160802 gel pcr 3.jpg|400px|thumb|right|Results]]
  
 
==== Sequencing of the gBlock 4.2 ====
 
==== Sequencing of the gBlock 4.2 ====
 
''By Caroline''
 
''By Caroline''
  
The PCR products obtain with Q5 on pPS16_008 were send to sequencing.
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The PCR products obtained with Q5 on pPS16_008 were sent for sequencing.
  
 
==== Culture of DH5alp|pPS16_003, DH5alp|pPS16_004, DH5alp|pPS16_006, DH5alp|pPS16_007 and DH5alp|pPS16_008 ====
 
==== Culture of DH5alp|pPS16_003, DH5alp|pPS16_004, DH5alp|pPS16_006, DH5alp|pPS16_007 and DH5alp|pPS16_008 ====

Latest revision as of 16:16, 9 October 2016

Tuesday 2nd August

Visualization

DreamTaq PCR on DH5alp|pPS16_001, DH5alp|pPS16_002, DH5alp|pPS16_005, DH5alp|pPS16_009, DH5alp|pPS16_0010, DH5alp|pPS16_011, DH5alp|pPS16_012, DH5alp|pPS16_013, DH5alp|pPS16_014, DH5alp|pPS16_015

By Charlène, Mathilde, Laetitia, Caroline and Léa

A Low fidelity PCR was carried out following the usual protocol and using the puc19 primers. 6 clones for each transformations were tested.

Each clone was spread and put at 37°C. We also did a liquid culture for each clone in 3mL of LB and Ampicilin(37°C and 180 rpm).

Results
Results
Results

Sequencing of the gBlock 4.2

By Caroline

The PCR products obtained with Q5 on pPS16_008 were sent for sequencing.

Culture of DH5alp|pPS16_003, DH5alp|pPS16_004, DH5alp|pPS16_006, DH5alp|pPS16_007 and DH5alp|pPS16_008

By Caroline

One colonie from each transformation was put into LB + Amp (50µg/mL) and incubated overnight at 37°C and 180rpm.

Interlab Study

Device 1 transformation

By Caroline

The device 1 was transformed in DH5alp following the usual Heat-shock protocol. The transformations were streaked on Petri dishes LB+Amp(50µg/mL)+IPTG(0.1µL/mL)+XGal(0.25µL/mL) and incubated overnight at 37°C.