DNA Reaction - Asymmetric PCR

Jeff Lawrence

1. Perform Standard PCR and purify a single band as Template DNA.

2. Dilute Template DNA to 1 pg/ mL.

3. Mix the following:

	1.0 µL	Template DNA 
	2.0 µL	10X TAQ Buffer (MgCl2 between 1 and 3 mM final)
	2.0 µL	8 mM dNTP (2 mM each)
	2.0 µL	5 mM Primer 1
	2.0 µL	0.5 mM Primer 2
	0.1 µL	TAQ Polymerase, at 5 unit/mL
	10.9 µL	ddH₂O						
	20.0 µL	Total volume

4. Cycle using the same reaction conditions as for a standard PCR reaction. The product will contain an excess of the ssDNA produced by Primer 1.

Last Update: Thursday June 19 2014

This page has been viewed Failed to execute CGI : Win32 Error Code = 2
times.

Eric Kofoid eckofoid at ucdavis.edu