Haynes Lab:Notebook/Jan/2016/06/15: Difference between revisions

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* Conclusions
* Conclusions
-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




Revision as of 18:46, 15 June 2016

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Genomic DNA Extraction, PCR, Agarose Gel Verification

  • 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. CMV = 588 		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


-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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