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
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Eric Kofoid
eckofoid at ucdavis.edu