Team:UNIPV-Pavia/Notebook/Week16

Week 16: 09/1/08 - 09/5/08
09/1/08
 * We had to perform these ligations:
 * Lig.22-R0062
 * Lig.31-Lig.22
 * but we didn't have enough Lig.22 and Lig.31 plasmids...


 * We infected 9 ml of LB + Amp with 30 µl of Lig.22 and Lig.31 glycerol stocks.

09/2/08
 * Glycerol stocks/miniprep for Lig.22 and Lig.31.


 * Plasmid digestion for:
 * Lig.22 (E-S)
 * Lig.22 (X-P)
 * Lig.31 (S-P)
 * R0062 (E-X)


 * Run/gel extraction.


 * Ligations:
 * Lig.31-Lig.22 (=Lig.35)
 * Lig.22-R0062 (=Lig.30)
 * We incubated ligations at 16°C overnight.

09/3/08
 * We transformed/plated ligations.

09/4/08
 * Colony PCR for the two ligation plates


 * Gel results:
 * Lig.30 (3rd colony)
 * Lig.35 (gel showed an unexpected contaminant...we decided to re-perform this ligation)

09/5/08
 * Glycerol stock/miniprep for Lig.30.


 * We sent Lig.30 (3rd col) to Primm for sequencing.