摘要
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.
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.
出处
《林业科学研究》
CSCD
北大核心
2003年第1期117-122,共6页
Forest Research
基金
2002-2004年浙江省自然科学基金项目"桤木属植物系统生物学及群体结构遗传研究"(编号301265)
