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, 19-Jun-2014 11:50:16 PDT
This page has been viewed 188 times.
Eric Kofoid eckofoid at ucdavis.edu