SBB11Ntbk-Helen Shi

Helen Shi 13:38, 17 February 2011 (EST)

Today, I got my PCR back and then I ran an analytical gel to check that the size of my PCR product is 531 basepairs. From the gel picture, my PCR product appears to be about 500 basepairs, which is correct. Thus, I continued on to the purification of my PCR product with a Zymo Cleanup. Tomorrow, I will digest my PCR product with EcoRI and BamHI.

I also digested the plasmid pBjk2741-jtk2936 with EcoRI and BamHI restriction enzymes to cut out the HlyE part. After an hour of digestion, I am performing a preparation gel to extract the digested HlyE part. Tomorrow, I may ligate and transform my HlyE part.

Helen Shi 13:44, 15 February 2011 (EST)

Today, I set up the PCR for my sbb1136 part using the prediluted oligos ss63r and ss63f (10 uM). Since the template for my part came from the MG1655 genome, I used the Expand buffer and polymerase for my PCR. I set up my PCR reaction with 1 uL each of my 10 uM oligos. From my construction file, my predicted PCR product has a size of 531 bp so the PCR file to be used is 2K55.

Helen Shi 04:29, 8 February 2011 (EST)

Hi!