High-throughput conformation-sensitive gel electrophoresis for discovery of SNPs
Leung, Y.F.; Tam, P.O.; Tong, W.C.; Baum, L.; Choy, K.W.; Lam, D.S.; Pang, C.P.
Biotechniques 30(2): 334-5 338-40
2001
ISSN/ISBN: 0736-6205 PMID: 11233602 Document Number: 535981
High-throughput screening for single nucleotide polymorphisms (SNPs) or mutations can be achieved by inexpensive technologies. We modified the original protocols of conformation-sensitive gel electrophoresis (CSGE) to increase throughput several fold to 1.3 samples/min, which is about five times faster than denaturing high-performance liquid chromatography (DHPLC). The modifications include decreasing the gel thickness, increasing the number of lanes to 96, and increasing the number of samples per lane to seven. This high-throughput CSGE method is fast, robust, and as simple as the original protocols. Together with a two-stage strategy for screening homozygotes and the replacement of ethidium bromide with SYBR(R) Gold DNA dye staining, this protocol is a reliable and cost-effective alternative for laboratories that require high-throughput screening.