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Magnetic bead purification of labeled DNA fragments for high-throughput capillary electrophoresis sequencing
1Joint Genome Institute/US Department of Energy, Walnut Creek, CA 94598, USA. elkin1@llnl.gov
Biotechniques
|June 21, 2002
Abstract:
We have developed an automated purification method for dye-terminator-based DNA sequencing products using a magnetic bead approach. This 384-well protocol generates sequence fragments that are essentially free of template DNA, salt, and excess dye-terminator products. In comparison with traditional ethanol precipitation protocols, this method uses no centrifugation, is rapid, completely automated, and increases the phred-20 read length by an average of 40 bases. To date, we have processed over 4 million samples with 94% averaging 641 phred-20 bases on the MegaBACE 1000 and 4000 and the ABI PRISM 3700 capillary instruments.