Long range PCRs for sequencing

rifat_h rifat_h at icr.ac.uk
Tue Nov 7 18:44:17 EST 1995


Hi,
	Has anyone tried long range PCR of fragments between 15kb-100bk and then sequenced
them either by subcloning or by direct sequencing?

If so what are the things to watch when doing long range PCRs. Also is there a danger of 
amplifying some other unknowns as part of your sequence. 

I am interested in doing a long range PCR to get a 15kb product and would be grateful if
anyone has any info on the best cycling times; primer design and primer template ratios.
Or any methods published and have been used successfully by the researcher.

Thanks in advance.

Rifat

rifat_h at icr.ac.uk




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