Friday 22^nd July

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Low Fidelity Dreamtaq PCR of DH5α|pPS16_002

By Mathilde

A DreamTaq PCR was made with DH5α|pPS16_002 cultures following the usual protocol with Tm at 57°c and 5min for the initial denaturation.

We divided up the PCR mix in 6 PCR tubes and added in each one a different clone from the transformed culture. The picked colonies were re-plated on a Petri dish with LB + Ampicillin (50µg/mL) + X-Gal/IPTG (1/1000).

Results : PCR products expected were :

The electropheresis on agarose gel showed absolutely no PCR products. pPS16_002 transformation from the 19/07/2016 is re-plated (50µL) on two Petri dishes LB + Ampicillin (50µg/mL) + X-Gal/IPTG (1/1000).

High Fidelity Q5 PCR of transformed DH5α with pPS16_004 and pPS16_007

By Laetitia

The PCR was performed following the usual protocol. 8 tubes were done: 5 clones of pPS16_004 and 3 clones of pPS16_007

The Tm was at 60°C

Glycerol stocks for DH5α transformed with pcl_TAA, pcl_TAG and pcl_Tq

By Laetitia

8 stocks were made: 2 clones (Cl 1 and Cl 2) :

• Cl1 and Cl2 of pcl_TAA
• Cl1 and Cl2 of pcl_TAG
• Cl1 and Cl2 of pcl_Tq

For 1 glycerol stock:

• 1mL of liquid culture
• 500 μL of glycerol 60%