Team:University of Ottawa/17 July 2008

(Difference between revisions)

Revision as of 14:46, 22 July 2008

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

@@ Line 14: / Line 14: @@
 ::<li> A transformation was performed on PTP2 ligation product - I then left the streaked plates to incubate for tomorrow.
 ::<li> A glycerol stock was finally made of the BY4642 yeast strain int. DQ232597.
+'''Dan'''
+:'''Gel of construct 1 after ligation'''
+::<li> Gel was unsucessful, the wrong primers were used for PCR amplification.
+::<li> Ligation seemed to work very efficiently, digesting in 50 uL with the new ClaI enzyme is working very well.
+:'''Digestions'''
+::<li> Three digestions were performed 1+T, 1+T (higher concentration of vector and insert), and just the vector
+:'''Ligations'''
+::<li>Three above plasmids were ligated overnight