Team:University of Ottawa/17 July 2008

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Today in the Lab
Chris
 * Purification of AtCRE
 * <li> used PCR clean up kit to purify AtCRE sample
 * Tranformation of Competent Cells
 * <li> followed transformation of competent cells protocol to insert AtCRE plasmid into preprepared competent E. coli cells.
 * <li> allowed cells to incubate overnight at 37 C

Matt
 * Ligation
 * <li> As suggested by Dan the ligation was spiked with 1 ul ATP.
 * <li> I then performed a PCR cleanup of the Ligation product in order to purify it for transformation into competent cells.
 * Transformation
 * <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

Tammy
 * Plasmid DNA Isolation
 * <li> XL10 Competent E.Coli cells transformed with pDR197::AtCKX2 were lysed and plasmid DNA was isolated using the Sigma-Aldrich kit.
 * <li> Average DNA concentration approximately 80 ng/&mu;L


 * Digestion of isolated pDR197::AtCKX2

<ol> <li> Control - No DNA (H2O) <li> PURE I <li> 1:10 II <li> 1:100 I <li> 1:100 II