Lengthening simple DNA repeats

ww40 William_D_WARREN at UMAIL.UMD.EDU
Thu Feb 24 14:49:00 EST 1994


Trevor Forsyth <pha23 at cc.keele.ac.uk> writes:

>Has anybody got any ideas for an efficent method of extending the length of
>a simple repetitive DNA? We currently have poly[d(c7A-T)].poly[d(c7A-T)]
>(i.e. the adenine is deaza at the 7 position) which from agarose is approx.
>1000 bp long and we wish to extend it to 10-20 k bp long. It has been
>suggested that I do some annealing, followed by Klenowing the gaps and then
>using T4 ligase to link everything together but not being a proper
>biochemist I am worried...
>
>Any advice?

Trevor,

You might like to read the following:

White et al., "Concatimer Chain Reaction: A taq DNA polymerase dediated
mechanism for generating long tandemly repetitive DNA sequences"
Anal. Biochem 199:184-190.

Rudert and Trucco "DNA polymers of protein binding sequences generated by PCR"
Nuc. Acids Res 18: 6460

Don't know how well Taq will handle 7deazadATP though.

Good luck
BILL

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William D. WARREN, PhD
Center for Agricultural Biotechnology     Email: ww40 at umail.umd.edu
University of Maryland at College Park    Phone: (301) 405-7681 

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