Lawrence Berkeley National Laboratory
Magnetic bead purification of labeled DNA fragments for high-throughput capillary
- Author(s): Elkin, Christopher
- Kapur, Hitesh
- Smith, Troy
- Humphries, David
- Pollard, Martin
- Hammon, Nancy
- Hawkins, Trevor
- et al.
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 ABI Prism 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.