IUBio

High throughput PCR purification

Barbara Simionati rhteam at eos.bio.unipd.it
Thu Nov 11 06:53:59 EST 1999


Hi,

We purify our pcr using 384-well plates (Polyflitronics glass fiber
filter plates) and a Multimek robot From Beckman (96 tips).
At the moment we mix the pcr (around 10 ul) with 3 volumes of  Qiagen PB
buffer (we are going to try other chaotropic agents), we transfere
averything to the filter plate and centrifuge, then we wash twice with
Qiagen PE buffer (or ethanol 80%) and elute with 20 ul of water.
We perform sequencing reactions and run them on a 3700 ABI sequencer and
we have very good sequences: the best ones have the first N 700-750 bp .


I would be interested in knowing the recipe of the Qiagen PB buffer if
someone has an idea of what could it be.

Best regards,

Barbara

Barbara Simionati
CRIBI Universita' di Padova
Dipartimento di Biologia
Complesso biologico
VIA G.Colombo, 3
35121 Padova, Italy
PHONE  +39-49-8276160
FAX    +39-49-8276159
E-MAIL rhteam at caos.bio.unipd.it






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