High-throughput conformation-sensitive gel electrophoresis for discovery of SNPs.
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 Gold DNA dye staining, this protocol is a reliable and cost-effective alternative for laboratories that require high-throughput screening.
Leung
Y F
YF
Chinese University of Hong Kong, Hong Kong.
Tam
P O
PO
Tong
W C
WC
Baum
L
L
Choy
K W
KW
Lam
D S
DS
Pang
C P
CP
D006579:Heterozygote; D006720:Homozygote; D020641:Polymorphism, Single Nucleotide