Haynes Lab:Notebook/Jan/2016/02/25

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PCR and Agarose Gel Electrophoresis on 4-5, 4-7, 4-9, and 6-4

  • Round 2: Genomic DNA prepped from candidate clonal lines
    • To be done after chosing a single primer pair per validation region
  1. BD4-5, Chromometer 2
  2. BD4-7, Chromometer 2
  3. BD4-9, Chromometer 2
  4. BD6-4, Chromometer 1


ROUND 2 PCR: Candidate clonal lines

Reagent Vol Mix* (x #) Expected:
1. CMV = 588
Hover name
5 μL/lane; 1% agarose; Ladder
genomic OR plasmid DNA 1.0 / 0.5 7.0 / 4.5
10 μM forward primer 1.0 ---
10 μM reverse primer 1.0 ---
2x GoTaq green 10.0 70.0
dH2O 7.0 / 7.5 49.0 / 52.5
  20.0 μL ---

Note: *Prep one Mix per template

  • For each template, aliquot 18.0 master mix into 6 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