Help! Separating multiple band PCRs for sequencing

May maya at ichr.uwa.edu.au
Wed Sep 23 22:34:21 EST 1998


Hi,
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.

May

-- 
maya at ichr.uwa.edu.au

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

Fax 61 9 388 3414



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