DNA Molecular Fingerprinting Construction Of Hordeum Vulgare By AFLP Marker

Barley (Hordeum Vulgare)is one of the most ancient crops in the world, mainly used in feed, grin, and industrial material for beer, medicine and health protection. The purity and quality of of barley is very importment for beer taste and quality. To set up a fast and accurate detection method for barley’s identification and selecting is very importment for beer industry. In this paper,27domestic and imported beer barley varieties were studied and the fingerprinting map were set up with AFLP method.It will be very useful for identification of barley varieties and beer industry. The main results as follows:1、We identifed the most suitable method for barley DNA extraction through comparison of different methods.The high-qulity barley genomic DNA extracted by the plant DNA genome Kit, which were completely suitable for the anaylsis of AFLP.2、AFLP analysis system suitable for barley genomic DNA was stablished:The purified barley genomic DNA(50ng/μL)5μL,EcoR I10U/μL0.5μL,Mse I10U/μL0.5μL,10×Tango buffer3μL, T4DNA ligase(5U/μL)0.2μL, were in a reaction volume of20μL and incubated at37℃and65℃for4h respectively. Double-stranded adaptor were added to the restriction firagment at22℃for10minutes. The pre-amplification products were diluted20times before selective amplification. And then it will start selective amplification.The selective amplification products were denatured by adding8μL Loading Buffer and heated up at95℃for5minutes. The PCR production were analysed by6%denatural aerylamide/bisacrylimade gel and the stained by silver.3、The primer combinations E+3/M+3can be used to amplify a proper number of bands and the AFLP fingerprinting was set up after the sliver-staining.3pairs primer from64pairs of primer combination were selected and abundant and clear bands, high polymorphismcan be got after AFLP amplify. And then we constructed AFLP fingerprinting of27beer barley varieties from domestic and overseas.