DNA concentrations for megaprimer annealing and extension

newsnet customer via methods%40net.bio.net (by customer At newsnet.com)
Thu Feb 1 06:35:40 EST 2007


> Dear experts,
>
> I would like to anneal two partially complementary pieces of DNA
> obtained by pcr  (about 500 bp each with a 50bp overlap) and extend
> them (adding dNTP and Pol). So each strand will compete for the
> complementary strand an the other strand with the overlapping region.
> Is there a concentration range where this might work best?
>
> Next step would be a PCR amplification with end-to-end primers.
>
> Your input is welcome!
>
> Best regards,
>
> WS

200ng for each primer. For your next step, use 5-10ng DNA.

ST 




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