Haynes Lab:Notebook/Jan/2016/06/21

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Redo of PCR/Gel Electrophoresis for 4-4, 4-5, 6-1, 6-2, 6-3, 6-4, 6-5, 6-6, 6-7 and Redo of Flow Cytometry Analysis on Synthetic Repressor Experiment

  • PCR: Candidate cell lines (3 primers + 1 error = 4)
Reagent Vol Mix* (x #) Expected:
1. 4-x, P1 = 1145
2. 4-x, P3 = 1046
3. 4-x, P6 = 1139
4. 6-x, P1 = 1146
5. 6-x, P3 = 1046
6. 6-x, P6 = 1146 		Hover name
Hover name
5 μL/lane; 1% agarose; Ladder
genomic OR plasmid DNA 1.0 / 0.5 4.0 / 2.0
10 μM forward primer 1.0 ---
10 μM reverse primer 1.0 ---
2x GoTaq green 10.0 40.0
dH2O 7.0 / 7.5 28.0 / 30
  20.0 μL 72.0 μL

Note: *Prep one Mix per template

  • For each template, aliquot 18.0 master mix into 3 tubes

Thermal cycler: Labnet - GOTAQ

  • 95°C 3 min.
  • 30x[95°C, 30 sec; 57°C 30 sec; 72°C 30 sec]
  • 72°C 3 min.
  • 4°C, ∞


  • Conclusions
  • Top image, top row, left to right: 4-4, 4-5, 6-1, 6-2, P1, 3, and 6 for each.
  • Top image, bottom row, 6-3, 6-4, 6-5, 6-6, P1, 3, and 6 for each.
  • Bottom image, top row, 6-7, P1, 3, and 6.
  • Bottom image samples on bottom row are something else.



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