quantitative PCR

Volker Blaschke vblasch at gwdg.de
Fri Jul 19 02:43:28 EST 1996


We do quantitative PCR with an internal standard derived from the 
original cDNA where a small fragment is removed by restriction enzyme 
digestion. Both can be coamplified in the same reaction mix and compete 
for the primers according to their molar ratio. If you take known 
amounts of your internal standard, you can determine the amount of 
wild type cDNA at the point where both PCR products are formed at equal 
amounts. Although be warned that this method is rather labour-intensive.

Dr. Volker Blaschke
Depts. of Biochemistry, Dermatology
Georg-August-University, Goettingen, Germany
Email: vblasch at gwdg.de
Tel: xx49 551 39 5959 / 5978

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