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Revision as of 14:55, 9 August 2016

Friday 29th July

Lab work

Visualization

gBlock 1.1, 1.2 and GFP1-9 insertion in puc19

By Caroline

The insertion was carried out following the usual protocol.

High fidelity PCR on bacteria transformed with pPS16_001, pPS16_002 and pPS16_005

By Alice and Mathilde

A PCR with Q5® High-Fidelity 2X Master Mix was performed on clones selected after screening PCR made on July 28, following this protocol. We choose clones which had the good size of insert. We selected the clone 2 for DH5α|pPS16_001; clones 4,7 and 8 for DH5α|pPS16_002 and clones 1 and 4 DH5α|pPS16_005. 1151_pheoR and 1152_pheoF primers were used. Annealing temperature was 66°C and initial denaturation was 3 min. After amplification, 1 µL of loading dye was added to 5µL of each PCR products. Then 5 µL of this mix were put into a well of the gel. A last well contained 10µL of DNA ladder. PCR products were migrated 30min at 100V.

PCR products expected were :

Plasmids Band size (bp)
pPS16_001 1017
pPS16_002 1017
pPS16_005 1017
File:T--Paris Saclay--160729 Visualisation PCR.jpeg
Migration of gBlocks PCR product





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