Primers fo genome amplification

Hemant Chikarmane hchikarmane at mediaone.net
Sun Jun 20 16:22:03 EST 1999


Juergen Oberstrass    <oberstra at hrz.uni-kassel.de> wrote:


>I want to clone unknown sequences from dictyostelium. Because I expect
>very few amount of material I want to amplify it before cloning by an
PCR
>step similar to DOP PCR of human material. Unfortunately dicty do not
have
>repetitive genomic elements between the coding sequences so that I
cannot
>use something like Alu-primers.

>...search or whatever operation on dicty sequences to find the most
abundant >twelve-mers and use this oligos for ramdom amplification. But
I have not >found any possibilitity to extract abundant sequences from
sequences >databases.

Dear Juergen,

Why don't you use RAPD-PCR primers (10-mers) to amplify Dictyo genomic
sequences, then you can clone these amplification products.  These
primers are commercially available (Operon, University of British
Columbia, etc.), so you wouldn't have to design your own.

Hemant Chikarmane
Marine Biological Laboratory
Woods Hole, MA  02543





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