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《Journal of Agricultural University of Hebei》 2005-03
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Establishment of SRAP technique system in wheat genome

WU Zhi-pu,YANG Wen-xiang,LIU Da-qun,ZHANG Ting (Department of Plant Pathology, Agricultural University of Hebei, Biological Control Center of Plant Pathogens and Plant Pests of Hebei Province, Baoding 071001, China)  
In this paper, some factors involving genomic DNA template, dNTP Mixture, Taq polymerase and PCR amplification time and temperatures of denature, annealing and amplification affecting the SRAP researching system of wheat were studied and an advanced technical system was established. The 20 μL reaction mixture contained: 25 ng of genomic DNA template, 188 mmol/L of dNTP, 0.75 U of Taq polymerase, 25 ng of primer and appropriate reaction buffer. Amplification was carried out in two steps: in the first 5 cycles, the annealing temperature was 37℃, and during the following 35 cycles, the annealing temperature was 50℃. In all the cycles, denaturation was carried out for 1 min at 94℃, annealing for 1 min, and amplification for 10 sec at 72℃. After all the cycles, amplification was done for 1 min at 72℃.
【Fund】: 国家自然科学基金(30170602)赞助项目
【CateGory Index】: S512.1
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