Error rate in Taq pol. 1:270?

Dr. Duncan Clark duncan at genesys.demon.co.uk
Tue Feb 11 04:49:52 EST 1997


In article <t.chappell-1002971432260001 at nntp-server.bcc.ac.uk>, Tom
Chappell <t.chappell at ucl.ac.uk> writes
>
>You're using faulty logic to calculate the error rate. If you've done a 30
>cycle PCR, the polymerase has synthesized more than 24,000 bases to
>generate the single DNA fragment that you've subcloned and sequenced. Your
>error rate would be about 1 in 8,000, which isn't too bad for Taq.


Unfortunately so are you.

30 cycles of exponential amplification will give 2 to the power of 30
fold synthesis ie 1,074,000,000 x 810bp. 

Error rates are 1 error every 10,000 bases suynthesised. You can't
simply divide the total bases synthesised by the error rate because
errors are copied from one cycle to another an extra errors added. 

For example if the target was 10000bp then after one cycle you have one
error. The next cycle copies that error and introduces another etc etc.



More information about the Methods mailing list