SBB11Ntbk-MaryWang: Difference between revisions
Line 34: | Line 34: | ||
0.5 Expand polymerase 1, | 0.5 Expand polymerase 1, | ||
0.5uL Template DNA | 0.5uL Template DNA | ||
Run at 2k55 |
Revision as of 13:01, 17 February 2011
Mary Wang 14:18, 17 February 2011 (EST)
Analytical gel run on part ssb1139: Used 2uL product from previous day and 5uL dye. Analytical gel shows that the part was done incorrectly.
REdone (x2)
One: 24uL ddH2O, 3.3uL 10X expand buffer, 3.3 uL dNTP, 1 uL ss45r, 1uL ss45f, 0.5uL template DNA, 0.5 expand polymerase 1
Two: 21.7uL ddH2O, 3.3uL DMSO, 3.3uL 10X expand buffer, 3.3uL dNTP, 1uLss45r, 1uL ss45f, 0.5uL template DNA, 0.5 expand polymerase 1
2k45 instead of 2k55
Digest pBjh1601KA-Bjh2296 using: uL of eluted PCR product, 1uL of NEB Buffer 2, 0.5uL EcoRI and 0.5uL BamHI
Mary Wang 14:16, 15 February 2011 (EST)
PCR ss45r/ss45f on MG1655 genome.
Used Expand polymerase because it was a genome.
Put template DNA into PCR tube first then the Expand polymerase second.
Did not dilute the oligos.
Used cloning by PCR- basic PCR method.
Amounts used: 24uL ddH2O, 3.3uL 10X expand buffer, 3.3uL dNTP, 1uL Oligo 1, 1uL Oligo 2, 0.5 Expand polymerase 1, 0.5uL Template DNA
Run at 2k55