Conversion of an RAPD marker to an STS marker for barley variety identification |
| |
Authors: | David Hoffman An Hang Steve Larson Berne Jones |
| |
Institution: | (1) Small Grain and Potato Germplasm Research Unit, USDA-ARS, 1691 S, 2700 W, 83210 Aberdeen, Idaho, USA;(2) Forage and Range Research Laboratory, USDA-ARS, Logan, Utah, USA;(3) Cereal Crops Research Unit, Barley & Malt Lab, USDA-ARS, Madison, Wisconsin, USA |
| |
Abstract: | Barley (Hordeum vulgare L.) variety identification is important to the malting and brewing industries. Because many new malting cultivars (varieties)
are closely related, new and more effective identification techniques are needed. We report on a series of techniques used
to convert an RAPD marker to a more stable STS marker that can identify barley Stander from Robust, an important distinction
for the American malting and brewing industries. The techniques included DNA extraction, RAPD amplification, random cloning
of all amplified fragments, selection of clones by insert size, DNA sequencing of select inserts, design of a barley-based
primer pair, and detection of a single nucleotide polymorphism using restriction endonucleaseAlu I. The barley-based primer pair was used to further sequence the RAPD fragment. Five single nucleotide polymorphisms between
Robust and Stander exist, one of which was detected by electrophoresing DNA fragments differentially restricted byAlu I. The conversion technique was different from ones previously reported in that it did not require manual extraction of DNA
fragments from a gel. This could be applied to other situations in which RAPD marker conversion would be desirable. |
| |
Keywords: | barley RAPD marker conversion SNP |
本文献已被 SpringerLink 等数据库收录! |
|