Magnetic bead purification of labeled DNA fragments for high-throughput capillary electrophoresis sequencing.
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.
Elkin
C
C
Joint Genome Institute/US Department of Energy, Walnut Creek, CA 94598, USA. elkin1@llnl.gov
Kapur
H
H
Smith
T
T
Humphries
D
D
Pollard
M
M
Hammon
N
N
Hawkins
T
T
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
D006018:Glycols; D014867:Water; D000431:Ethanol; D004247:DNA