SBB11Ntbk-MaryWang: Difference between revisions
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Used 2uL product from previous day and 5uL dye. | Used 2uL product from previous day and 5uL dye. | ||
Analytical gel shows that the part was done incorrectly. | Analytical gel shows that the part was done incorrectly. | ||
Analytical Gel2- lane 5. | |||
REdone (x2) | REdone (x2) | ||
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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 | 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 | Run at 2k45 instead of 2k55 | ||
Digest pBjh1601KA-Bjh2296 using: | Digest pBjh1601KA-Bjh2296 using: | ||
8uL of eluted PCR product, 1uL of NEB Buffer 2, 0.5uL EcoRI and 0.5uL BamHI. | |||
Prep Gel run- shown on PrepGel2- lane 4. | |||
Cut out ~837 part- melted with 600uL ADB buffer then froze. | |||
==[[User:Mary Wang|Mary Wang]] 14:16, 15 February 2011 (EST)== | ==[[User:Mary Wang|Mary Wang]] 14:16, 15 February 2011 (EST)== | ||
Revision as of 13:17, 18 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. Analytical Gel2- lane 5.
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
Run at 2k45 instead of 2k55
Digest pBjh1601KA-Bjh2296 using: 8uL of eluted PCR product, 1uL of NEB Buffer 2, 0.5uL EcoRI and 0.5uL BamHI. Prep Gel run- shown on PrepGel2- lane 4. Cut out ~837 part- melted with 600uL ADB buffer then froze.
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
