A Low‐Cost, High‐Throughput Polyacrylamide Gel Electrophoresis System for Genotyping with Microsatellite DNA Markers
- 1 September 2003
- journal article
- research article
- Published by Wiley in Crop Science
- Vol. 43 (5) , 1828-1832
Abstract
Microsatellite DNA markers are widely used in genetic research. Their use, however, can be costly and throughput is sometimes limited. The objective of this paper is to introduce a simple, low‐cost, high‐throughput system that detects amplification products from microsatellite markers by nondenaturing polyacrylamide gel electrophoresis. This system is capable of separating DNA fragments that differ by as little as two base pairs. The electrophoresis unit holds two vertical 100‐sample gels allowing standards and samples from a 96‐well plate to be analyzed on a single gel. DNA samples are stained during electrophoresis by ethidium bromide in the running buffer. In addition, one of the gel plates is UV‐transparent so that gels can be photographed immediately after electrophoresis without disassembling the gel‐plate sandwich. Electrophoresis runs are generally less than two hours. The cost per gel, excluding PCR cost, is currently estimated at about $2.60, or less than $0.03 per data point. This system has been used successfully with soybean [Glycine max (L.) Merr.] and wheat (Triticum aestivum L.) microsatellite markers and could be a valuable tool for researchers employing markers in other species.Keywords
This publication has 7 references indexed in Scilit:
- AFLP in Triticum aestivum L.: patterns of genetic diversity and genome distributionEuphytica, 2002
- Loci underlying resistance to Race 3 of soybean cyst nematode in Glycine soja plant introduction 468916Theoretical and Applied Genetics, 2001
- Detection of errors in dinucleotide repeat typing by nondenaturing electrophoresisElectrophoresis, 2001
- Mapping and genome organization of microsatellite sequences in rice (Oryza sativa L.)Theoretical and Applied Genetics, 2000
- The physical mapping of microsatellite markers in wheatGenome, 1998
- Molecular mapping of wheat. Homoeologous group 3Genome, 1995
- Molecular-genetic maps for group 1 chromosomes of Triticeae species and their relation to chromosomes in rice and oatGenome, 1995