Haynes Lab:Notebook/Short Projects/2015/04/09

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04/09/15 - Alexander Ellingson

PCR-verify activation domain primers
Try different cDNA libraries for the CARM1 and MYB fragment to account for any mutations or deletions. Last 2 fragments run to verify for any variations. iPS cell template utilized in place of U20S, SKNSH, and K562 templates.

  1. iPS, 1:100 cDNA dilution; SMCA4 = 4428 bp: SMCA4 F1/SMCA4 R1; ANM5 = 1911 bp: ANM5 F1/ ANM5 R1.


Reagent Rxn1 Rxn2 Rxn3 Expected:
1. SMCA4 = 4428 bp
2. ANM5 = 1911 bp: ANM5 F1/ ANM5 R1;. 		Hover name
10 μL/lane, 1% agarose; Ladder
Template 2.0 2.0 2.0
10 uM fwd primer 1.0 1.0 1.0
10 uM rev primer 1.0 1.0 1.0
2x GoTaq green 12.5 12.5 12.5
dH2O 9.5 9.5 9.5
  25.0 25.0 25.0

Program: GOTAQ35cyc

  • 95°C, 3 min
  • 35x[95°C, 30 sec; 57°C, 30 sec; 72°C, 2 min]
  • 72°C, 5 min
  • 4°C ∞










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