Gewtting the concentration of PCR product and cloning

[Aawara Chowdhury]

In <mailman.1235.1230408039.29717.methods from net.bio.net>,
 shifali  chatrath <shifalich from rediffmail.com> wrote:

>  ?
>
> Dear all!
> Please read carefully what DK wrote before pointing fingers at me!
> Mr. Aawara is also repeating the same thing.[PCR products amplified
> with a proof-reading polymerase must be A-tailed ]. Please note that, if the enzyme has proof reading activity, it will remove 3'A and TA cloning cannot be done. And, it is Taq's property to put an extra A at 3' which has nothing to do with its proof reading.

The problem is with your comprehension of English.  Let me clarify:

If one wrote, "PCR products .... ARE A-tailed", then it would indicate
that products made with a proof-reading polymerase are already A-tailed.

On the other hand, if one writes "PCR products .... MUST BE A-tailed", then
it indicates that the products made with a proof-reading polymerase are
NOT A-tailed, and therefore MUST BE A-tailed for them to be cloned into a 
TA-vector.

Hope that helps you .....

AC
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