User:Madeline Hartman/Notebook/Dr. Hartings Lab/2012/11/06: Difference between revisions
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I ran two PCR reactions with the following content: | I ran two PCR reactions with the following content: | ||
Tube 1- | |||
# 40.6 μL distilled H2O | # 40.6 μL distilled H2O | ||
# 5.0 μL 10x cloned Pfu buffer | # 5.0 μL 10x cloned Pfu buffer | ||
Line 16: | Line 17: | ||
# 1.0 μL Primer 2 (ng/μL) | # 1.0 μL Primer 2 (ng/μL) | ||
# 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/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: | |||
[[Image:MH 20121106Table of cycles.png|300px]] | |||
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 | |||
Revision as of 10:00, 4 December 2012
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Entry titleI ran two PCR reactions with the following content: Tube 1-
and Tube 2-
Tube 3-
Tubes 2 and 3 were later combined. Primer 1- t109c forward Primer 2- t109c reverse The three tubes were ran for the following cycles:
Tubes were left in thermocycler overnight at 4°C
|