PCR Amplification of DNA: Difference between revisions

Reddj (talk | contribs)
updated
Reddj (talk | contribs)
updated
Line 9: Line 9:
==Protocol==
==Protocol==
*Use the following volumes per reaction
*Use the following volumes per reaction
::*Buffer, 5 uL of 10X buffer (Dave's fridge)
::*12.5 uL DreamTaq
::*Primers, 10uL of 1uM stock solution in water (both primers combined)
::*7.5 uL RNAse-Free water
::*dNTPs, 5uL of 2 mM ("molecular biology stuff" box in freezer)
::*5.0 uL Working stock Primer (of 0.4-1uM stock solution in water (both primers combined))
::*Sterile water, 28 uL
 
::*Template 1 uL
==PCR Program==
::*Polymerase 1 uL (turbo pfu found in "enzymes" box in freezer)
*For animals: use program denoted in [[Genotyping Program]]


*Run PCR Program (approx 3.5 to 4 hours).  Normally use touchdown PCR ('''DAVETD''') as follows:
*Run PCR Program (approx 3.5 to 4 hours).  Normally use touchdown PCR ('''DAVETD''') as follows: