摘要
[Objective] The research aimed to screen out optimum RAPD reaction system on genomic DNA of Agrocybe chaxingu Huang.[Method] The single factor experiment was adopted to select the required Mg2+ concentration, template DNA concentration,primer concentration,dNTPs concentration,Taq enzyme concentration and anneal temperature initially.[Result] The optimum reaction system for RAPD amplification of Agrocybe chaxingu Huang was listed as follows:2.5 μl Buffer, 2 mmol/L Mg2+, 75 ng DNA, 0.5 μmol/L primer, 150 μmol/L dNTPs and 2.0 Taq enzyme.The reaction process was also listed as follows: denaturation for 5 min at 92 ℃,35 cycles(1 min at 92 ℃, 1 min for 35.5 ℃ and elongation for 2 min at 72 ℃),10 min at 72 ℃.[Conclusion] The research provided reference for conducting RAPD analysis of and studying genetic relationship and genetic diversity of Agrocybe chaxingu Huang.
[Objective] The research aimed to screen out optimum RAPD reaction system on genomic DNA of Agrocybe chaxingu Huang.[Method] The single factor experiment was adopted to select the required Mg2+ concentration, template DNA concentration,primer concentration,dNTPs concentration,Taq enzyme concentration and anneal temperature initially.[Result] The optimum reaction system for RAPD amplification of Agrocybe chaxingu Huang was listed as follows:2.5 μl Buffer, 2 mmol/L Mg2+, 75 ng DNA, 0.5 μmol/L primer, 150 μmol/L dNTPs and 2.0 Taq enzyme.The reaction process was also listed as follows: denaturation for 5 min at 92 ℃,35 cycles(1 min at 92 ℃, 1 min for 35.5 ℃ and elongation for 2 min at 72 ℃),10 min at 72 ℃.[Conclusion] The research provided reference for conducting RAPD analysis of and studying genetic relationship and genetic diversity of Agrocybe chaxingu Huang.
基金
北京市科委重点项目(D0705007040291)~~
