Taking China’s endemic tree species Alnus cremastogyne and other 5 Alnus species as test materials, the DNA extraction of genome and the amplification of PCR were studied. Two methods were tested, i.e. freezi...Taking China’s endemic tree species Alnus cremastogyne and other 5 Alnus species as test materials, the DNA extraction of genome and the amplification of PCR were studied. Two methods were tested, i.e. freezing treatment+5% CTAB and liquid nityrogen+5% CTAB. It is proved that the optimum PCR program is as follows: predenaturating under 94 ℃ for 3 min., denaturating under 94 ℃ for 18 seconds, annealing under 36 ℃ for 80 seconds and extending under 72 ℃ for 120 seconds. After 40 cycles, the sample is reacted for 10 minutes under 72 ℃. PCR system includes buffer 2.5 μL, dNTP 2.5 μL, primer (s60+s155) 2 mmol·L-1, Mg2+ 3.0 mmol·L-1, Tap enzyme 1 U, DNA 40 mg, then adding ddH2O to 20 μL. This study may provide some references for the application of RAPD technique in genetic research of alder tree species.展开更多
文摘Taking China’s endemic tree species Alnus cremastogyne and other 5 Alnus species as test materials, the DNA extraction of genome and the amplification of PCR were studied. Two methods were tested, i.e. freezing treatment+5% CTAB and liquid nityrogen+5% CTAB. It is proved that the optimum PCR program is as follows: predenaturating under 94 ℃ for 3 min., denaturating under 94 ℃ for 18 seconds, annealing under 36 ℃ for 80 seconds and extending under 72 ℃ for 120 seconds. After 40 cycles, the sample is reacted for 10 minutes under 72 ℃. PCR system includes buffer 2.5 μL, dNTP 2.5 μL, primer (s60+s155) 2 mmol·L-1, Mg2+ 3.0 mmol·L-1, Tap enzyme 1 U, DNA 40 mg, then adding ddH2O to 20 μL. This study may provide some references for the application of RAPD technique in genetic research of alder tree species.
