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_h at icr.ac.uk