Magnetic bead purification of labeled DNA fragments forhigh-throughput capillary electrophoresis sequencing
We have developed an automated purification method for terminator sequencing products based on a magnetic bead technology. This 384-well protocol generates labeled DNA fragments that are essentially free of contaminates for less than $0.005 per reaction. In comparison to laborious ethanol precipitation protocols, this method increases the phred20 read length by forty bases with various DNA templates such as PCR fragments, Plasmids, Cosmids and RCA products. Our method eliminates centrifugation and is compatible with both the MegaBACE 1000 and ABIPrism 3700 capillary instruments. As of September 2001, this method has produced over 1.6 million samples with 93 percent averaging 620 phred20 bases as part of Joint Genome Institutes Production Process.
- Research Organization:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Organization:
- USDOE Director. Office of Science. Office of Biological andEnvironmental Research, Lawrence Livermore National Laboratory, LosAlamos National Laboratory
- DOE Contract Number:
- DE-AC02-05CH11231
- OSTI ID:
- 860368
- Report Number(s):
- LBNL-48920; BTNQDO; R&D Project: 625909; BnR: KP1103010; TRN: US200524%%117
- Journal Information:
- Biotechniques, Vol. 32, Issue 6; Related Information: Journal Publication Date: June 2002; ISSN 0736-6205
- Country of Publication:
- United States
- Language:
- English
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