Help! Separating multiple band PCRs for sequencing

May maya at
Wed Sep 23 22:34:21 EST 1998

I've been trying to separate out multiple bands obtained by PCR.

Bands were obtained after subtractive hybridisation and all products
contain artificially added adaptor sequences, which is why I ended up with
so many bands.

I've managed to reduce the products down to 2-4 bands per reaction from
about 25 bands by using BAND STAB PCR (where bands run on a gel are simply
stabbed by a pipette tip that is then placed in a PCR mix - that provides
sufficient template for a PCR to proceed).

However, I haven't been able to achieve complete separation.  Is anyone
out there aware of any methods that I can use other than using a
sequencing gel or cloning?

Your help would be greatly appreciated.


maya at

TVW Telethon Institute for Child Health Research
(Company Limited by Guarantee)
Western Australia

Fax 61 9 388 3414

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