Haynes Lab:Notebook/Short Projects/2015/03/11

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PCR-verify activation domain primers
Try different cDNA libraries for the CARM1 and MYB fragment to account for any mutations or deletions

  1. U2OS C002, 1:1000 cDNA dilution; KMT2A = 1134 bp: KMT2A F1/ KMT2A R1; KMT2C = 423 bp: KMT2C F1/ KMT2C R1; KMT2D = 423 bp: KMT2D F1/ KMT2D R1.
  2. SKNSH C001, 1:1000 cDNA dilution; same.
  3. K562 C001, 1:1000 cDNA dilution; same.


Reagent Rxn1 Rxn2 Rxn3 Expected:
1. U2OS KMT2A = 1134 bp
2. SKNSH KMT2A = 1134 bp
3. K562 KMT2A = 1134 bp
4. U2OS KMT2C = 423 bp
5. SKNSH KMT2C = 423 bp
6. K562 KMT2C = 423 bp
7. U2OS KMT2D = 423 bp
8. SKNSH KMT2D = 423 bp
9. K562 KMT2D = 423 bp
Hover name
10 μL/lane, 1% agarose; Ladder
Template 1.0 1.0 1.0
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 ∞


  • primers 4-6 attempted in order to rule out the possibility of primer discrepancy; the camera is damaged, so manual imaging techniques are necessary.







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