Indian Journal of Science and Technology

Abstract

DNA marker technology represents a potential tool for molecular breeding. The successful deployment of this technology depends on availability of reliable and simple genotyping platforms. But, most of the lab which dealing with molecular breeding has poor lab facility and they rely on horizontal agarose gel electrophoresis for genotyping. The poor resolution provided by this method in terms of allele differences is the major setback for research community. Therefore, we tried to introduce a simple method for upgrading the existing horizontal agarose units into horizontal polyacrylamide gel electrophoresis (H-PAGE) units with few amendments. This method can provide higher resolution with less effort and allows easy genotyping for different markers systems as compared to vertical PAGE units. In-order to reduce time, we employed fast staining methods such as ethidium bromide and fast silver staining for 20 min. The performance of modified unit was checked by separating and scoring SSR, SRAP and TRAP markers on 30 pigeon pea genotypes. Number of bands scored and size range observed for each marker technique was comparable to earlier results as reported for vertical PAGE based genotyping. Therefore, these H-PAGE units could be a valuable tool for molecular breeding.

Keywords: Horizontal Polyacrylamide Gel, Marker Genotyping