Big Dye v3.0 Dilutions?

Mark T Jung MARK.T.JUNG at USA.dupont.com
Fri Dec 21 08:03:35 EST 2001


Dennis,

We have tested v3.0 at 1/4 strength for PCR product sequencing (2ul BigDye,
6ul buffer, 7.5ng DNA per 100bp product, 20ul reaction).  This works at
least as well as v2.0 at 1/4 strength (as defined by the number of phred20
basecalls) and in some cases a little better.  We have not tested it yet at
higher dilutions.

Mark Jung




Dennis Shubitowski <shubitow at cvm.msu.edu>@hgmp.mrc.ac.uk on 12/20/2001
03:51:34 PM

Sent by:    owner-autoseq at hgmp.mrc.ac.uk


To:    autoseq at net.bio.net
cc:
Subject:    Big Dye v3.0 Dilutions?


Is anyone running diluted protocols using the Big Dye 3.0 kit? I am doing
small scale PCR based sequencing and have only seen protocols for earlier
versions of diltutions for the Big Dye chemistries. The 1X protocol seems
like a large waste of terminator for the price.

Thank you!

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