Team:UNIPV-Pavia/Notebook/Week16
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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.