ABI 3100: red dye blob at 240 basecalls

Phillip San Miguel pmiguel at purdue.edu
Tue Jul 3 08:11:10 EST 2001

Roland.Varecka at vie.boehringer-ingelheim.com wrote:

> Dear Angela, dear Steve,
> thanks for your hints on loading the multiscreen plates. I ran some test
> yesterday
> and the trend is clear: either reducing the loading volume from 30uL to 15uL
> (plus making sure the samples are applied exactly at center column) or increasing
> the amount of Sephadex G-50 from 45uL to 90uL produced very nice and clean sequences.
> The latter method gave more consistent sample quality and samples are more
> easily loaded when using an 8-channel pipettor, however may be a bit messy for routine application,
> as the swollen gel slightly protrudes over the rim of the wells (reminded me
> a bit of freshly tapped draft beers). I have also ordered the Quiagen DyeEx
> 96 kit.
> [...]

Actually, of the 3 types of commercial clean-up plates we have used: Qiagen, BioRad and Edge; Qiagen uses the least
amount of column material. In our hands BioRad and Edge gave superior results. Edge, especially seemed to be able to
completely remove large amounts of unincorporated dye-terminators.

Phillip SanMiguel
Purdue Genomics Core Facility


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