Magnetic Bead Purification of Labeled DNA Fragments for High-Throughput Capillary Electrophoresis Sequencing. (June 2002)

• Record Type: Journal Article
• Authors: Elkin, C.; Kapur, H.; Smith, T.; Humphries, D.; Pollard, M.; Hammon, N.; Hawkins, T.
• Is Part Of: Biotechniques. Volume 32:Number 6(2002)
• Page Start: 1296
• Page End: 1302
• Publication Date: 2002-06
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/02326st05
• Languages: English
• ISSNs: 0736-6205

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 TM 1000 and 4000 and the ABI PRISM ® 3700 capillary instruments.