Magnetic bead purification of labeled DNA fragments for high-throughput capillary electrophoresis sequencing

Biotechniques. 2002 Jun;32(6):1296, 1298-1300, 1302. doi: 10.2144/02326st05.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA / isolation & purification*
  • Electrophoresis, Capillary / methods*
  • Ethanol / metabolism
  • Glycols / metabolism
  • Magnetics
  • Microspheres
  • Water / metabolism

Substances

  • Glycols
  • Water
  • Ethanol
  • DNA