Line 61:
Line 61: ===Interlab Study===
===Interlab Study===
− ==== ====
+ ==== Preculture of device one transformed DH5a ====
− ''By ''
+ ''By Lea ''
+ Two colonies were inoculated inside of two tubes of 3mL of liquid LB medium containing 30 µg/ml Chloramphenicol. The tubes were incubated at 37°C overnight.
{{Team:Paris_Saclay/notebook_footer}}
{{Team:Paris_Saclay/notebook_footer}}
Revision as of 14:33, 4 August 2016
Tuesday 3st August Lab work Visualization Glycerol stocks By Caroline
The bacteria transformed with the plasmids sent the 2/08/2016 and those that would be sent the 4/08/2016 were put into glycerol. 1mL of liquid culture and 0.5mL of glycerol were put at -20°C.
High fidelity Q5 PCR on DH5alp|pPS16_004, DH5alp|pPS16_006 and DH5alp|pPS16_007 By Caroline
Q5 PCR was carried out following the usual protocol adapted to 50µL at a TM of 72°C. The primers used were specific to amplify only the interested sequence.
Extraction of the plasmids containing gBlocks pPS16_001, pPS16_002, pPS16_003, FRB, sg-ST1, FKPB, DS-NMcasN-, spacer and detection By Laetitia
The extraction was performed without kit, following the usual protocol .
It was done on:
Puc 19 containing
Clone
pPS16_001
4
pPS16_002
3
pPS16_003
2, 5
FRB
5
sg-ST1
2, 3, 5
DS-NMcasN-
3
FKBP
4
Spacer
1, 6
Detection
3, 4, 6
By Naiane
The kit "Charge Switch-Pro Plasmid Miniprep" was used to extract plasmidic DNA of the plasmid pPS16_003 containing the Gblock 2.1, plasmid pPS16_004 containing the Gblock 2.2, plasmid pPS16_006 containing the Gblock 3.2 and the plasmid pPS16_007 containing the Gblock 4.1 from 3mL of overnight culture.
Plasmids were resuspended in 100μL of Milli-Q water.
DNA stored at -20°C.
Interlab Study Preculture of device one transformed DH5a By Lea
Two colonies were inoculated inside of two tubes of 3mL of liquid LB medium containing 30 µg/ml Chloramphenicol. The tubes were incubated at 37°C overnight.
