PCR Design

Duncan Clark via methods%40net.bio.net (by blackhole from abuse.plus.com)
Wed Oct 11 03:30:11 EST 2006


Historians believe that in newspost 
<diLWg.22242$vi3.640 from bignews3.bellsouth.net> on Tue, 10 Oct 2006, Aawara 
Chowdhury <aawara from FEMA-trailer.org> penned the following literary 
masterpiece:
>> Speaking of which: What is expected % of PCR products with added Ts
>> overgangs when using Taq? The question recently poped up discussing
>> a particular experiment and I relized I have no idea. So, what is the
>> frequency? 10%? 50%? Nearly 100%?
>>
>
>The propensity of Taq to create products with a single base non-templated
>3' dT overhang is nil.  But close to 100% of the products will have a single
>base non-templated 3' dA overhang.

But is also very dependant upon the 5' sequence of the primer according 
to an 'old' genotyping Biotechniques paper. By altering the primer 
sequence they could ensure that 100% of products had the overhang and 
defined a the required sequence to achieve this. Not sure of the exact 
reference though :-(

Duncan
-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.


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