PCR Design

Jose de las Heras via methods%40net.bio.net (by josenet from tiscali.co.uk)
Tue Oct 10 19:42:42 EST 2006


"Aawara Chowdhury" <aawara from FEMA-trailer.org> wrote in message 
news:diLWg.22242$vi3.640 from bignews3.bellsouth.net...
> In <tkyWg.529$Mr7.4 from newsfe04.lga>,
> DK <dk from no.email.thankstospam.net> wrote:
>
>> In article <4ovkkpFgddqjU1 from individual.net>, "Jose de las Heras" 
>> <josenet from tiscali.co.uk> wrote:
>>>
>>>although an intermediate cloning in something like pGEM-T makes things
>>>easy... assuming the polymerase you're using adds Ts at high frequency.
>>
>> 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.
>
> AC

ah, well spotted!!!  :-)

it's A overhangs, guys... not T... the T is in the pGEM-T Easy vector, of 
course!
                                                                             
    ^^^

Jose 




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