Team:University of Ottawa/17 July 2008
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Today in the Lab
Chris
- Purification of AtCRE
- used PCR clean up kit to purify AtCRE sample
- Tranformation of Competent Cells
- followed transformation of competent cells protocol to insert AtCRE plasmid into preprepared competent E. coli cells.
- allowed cells to incubate overnight at 37 C
Matt
- Ligation
- As suggested by Dan the ligation was spiked with 1 ul ATP.
- I then performed a PCR cleanup of the Ligation product in order to purify it for transformation into competent cells.
- Transformation
- A transformation was performed on PTP2 ligation product - I then left the streaked plates to incubate for tomorrow.
- A glycerol stock was finally made of the BY4642 yeast strain int. DQ232597.
Dan
- Gel of construct 1 after ligation
- Gel was unsucessful, the wrong primers were used for PCR amplification.
- Ligation seemed to work very efficiently, digesting in 50 uL with the new ClaI enzyme is working very well.
- Digestions
- Three digestions were performed 1+T, 1+T (higher concentration of vector and insert), and just the vector
- Ligations
- Three above plasmids were ligated overnight