Haynes Lab:Notebook/Jan/2016/06/15

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Genomic DNA Extraction, PCR, Agarose Gel Verification, Freezer Inventory

  • Finished extracting genomic DNA from 4-1, 4-2, 4-3, 6-1, 6-2, 6-5, 6-6.

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
15 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

On the top, left to right: 4-1, 4-2, 4-3, P1, 3, and 6 for each. On the bottom, left to right: 6-1, 6-2, 6-5, 6-6, P1, 3, and 6 for each.


  • -150 Freezer Inventory, Box B6:
  • No 4-1 vials (one active T75 flask)
  • No 4-2 vials (one active T75 flask)
  • One 4-3 vial
  • One 4-4 vial
  • Three 4-5 vials
  • Two 4-6 vials
  • No 4-7 vials (No active flasks, gone forever)
  • No 4-8 vials (No active flasks, gone forever)
  • Two 4-9 vials


  • No 6-1 vials (one active T75 flask)
  • One 6-2 vial
  • Three 6-3 vials
  • No 6-4 vials (No active flasks, gone forever)
  • One 6-5 vial
  • One 6-6 vial
  • One 6-7 vial


  • Two U2-OS wild-type vials



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