Plant & Animal Genome IX Conference Plant & Animal Genome IX Conference
Poster: Sequencing & EST
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Successful high-throughput DNA sequence or SNP discovery analyses require that sequencing reaction purification be reliable, rapid and automatable. Common methods for purifying sequencing reactions involve several steps and require processes that are not easily automated. Prolinx,® Inc. has developed the unique RapXtract Dye Terminator Removal Kit that is rapid, easy to perform, yields high quality product and does not require flammable solvents or modified primers. This system uses a magnetic particle format and removes dye terminator artifacts by mass action. The protocol has three steps: 1. Remove storage buffer from particles, 2. Add sequencing reactions and mix, and 3. Remove purified extension products. The mixing step can be performed by vortexing or by pipette agitation, making it amenable to any automation system. Samples purified with RapXtract kits yield sequences with good Phred quality scores on the commonly used sequencing instruments and on the new ABI PRISM 3100. The kit has been optimized to minimize the use of expensive sequencing reagents and can be performed on reactions containing as little as 1īl of Big Dye reaction mix. Dye primer reactions and other dye terminator chemistries can also be purified with this system. The RapXtract kit has been successfully used to purify sequencing reactions from various DNA templates, including those from plant or animal sources. In addition, the RapXtract Dye Terminator Removal Kit has been used for analysis of polymorphisms in clinical samples.