Entry title
I ran two PCR reactions with the following content:
Tube 1-
- 40.6 μL distilled H2O
- 5.0 μL 10x cloned Pfu buffer
- 0.4 μL dNTPs (25 mM each)
- 1.0 μL DNA template (100 ng/μL)
- 1.0 μL Primer 1 (ng/μL)
- 1.0 μL Primer 2 (ng/μL)
- 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)
and
Tube 2-
- 20.3 μL distilled H2O
- 5.0 μL 10x cloned Pfu buffer
- 0.4 μL dNTPs (25 mM each)
- 1.0 μL DNA template (100 ng/μL)
- 1.0 μL Primer 1 (ng/μL)
- 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)
Tube 3-
- 20.3 μL distilled H2O
- 5.0 μL 10x cloned Pfu buffer
- 0.4 μL dNTPs (25 mM each)
- 1.0 μL DNA template (100 ng/μL)
- 1.0 μL Primer 2 (ng/μL)
- 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)
Tubes 2 and 3 were later combined.
Primer 1- t109c forward
Primer 2- t109c reverse
The three tubes were ran for the following cycles:
After 5 cycles, tubes 2 and 3 were combined, and run the rest of the cycles together.
Tubes were left in thermocycler overnight at 4°C
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