PCR Design

Jose de las Heras josenet at tiscali.co.uk
Mon Oct 9 18:06:05 EST 2006


"DK" <dk at no.email.thankstospam.net> wrote in message 
news:tkyWg.529$Mr7.4 at newsfe04.lga...
> In article <4ovkkpFgddqjU1 at individual.net>, "Jose de las Heras" 
> <josenet at 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%?
>
> Thanks,
>
> DK

My answer will be most unhelpful... because it's so vague, but here it is 
;-)

I seem to recall that a while ago I came across some info regarding the 
polymerase I was using (which I forget what it was) and I think there was 
some mention of anything between 50-75% fragments ocntaining a T overhang at 
each end.

Not really helpful, I know... :-(

Jose 




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