title: Magnetic bead purification of labeled DNA fragments for high-throughput capillary electrophoresis sequencing.
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.
authors.affiliation: Joint Genome Institute/US Department of Energy, Walnut Creek, CA 94598, USA. elkin1@llnl.gov
authors.forename: C
authors.initials: C
authors.lastname: Elkin
authors.forename: H
authors.initials: H
authors.lastname: Kapur
authors.forename: T
authors.initials: T
authors.lastname: Smith
authors.forename: D
authors.initials: D
authors.lastname: Humphries
authors.forename: M
authors.initials: M
authors.lastname: Pollard
authors.forename: N
authors.initials: N
authors.lastname: Hammon
authors.forename: T
authors.initials: T
authors.lastname: Hawkins
mesh_terms: D004247:DNA / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary / Q000379:methods*; D000431:Ethanol / Q000378:metabolism; D006018:Glycols / Q000378:metabolism; D008280:Magnetics; D008863:Microspheres; D014867:Water / Q000378:metabolism
chemical_list: D006018:Glycols; D014867:Water; D000431:Ethanol; D004247:DNA