User:Daniel A. Vargas/Notebook/General lab notebook/2015/07/07

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Gel electrophoresis of PCR amp

Reagent Volume Expected:
1. ANM5 = 1911,
2. MYB = 159
3. Carm1 = 1827
4. SETD7 = 459
5. KMT2E = 1176
6. SETD1A = 420
File:0707danielpcr.jpeg
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 1:100 1.0 μL
1X GoTaq 12.5
F primer 1.0
R Primer 1.0
dH2O 9.5
  25 μL

Program: Gotaq35cyclo (block A lane5)

  • 95°C, 3 min
  • 35x[95°C, 10 sec; 48°C, 30 sec; 72°C, 120 sec]
  • 72°C, 5 min
  • 4°C ∞

Program: Gotaq35cyclo (block B 1-4, and 6)

  • 95°C, 3 min
  • 35x[95°C, 10 sec; 64°C, 30 sec; 72°C, 120 sec]
  • 72°C, 5 min
  • 4°C ∞


Conclusion

Bands not able to be seen well enough, need to use a higher concentration. Dilution was 250ng/μL to a 2.5ng/μL concentration, this was advised by a fellow lab member. I will try and use a 1:10 to hit a perfect middle ground so bands will be clearly visible.