文献类型：期刊文献

中文题名：桤木群体遗传分化研究I.DNA提取和PCR条件的建立

英文题名：Research on the Populations Variation of Alnus cremastogyneI. DNA Extracting and Protocol Optimum for PCR

作者：卓仁英[1] 孟现东[1] 陈益泰[1]

第一作者：卓仁英

机构：[1]中国林业科学研究院亚热带林业研究所

年份：2003

卷号：16

期号：1

起止页码：117-122

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

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)

语种：中文

中文关键词：群体遗传分化;桤木;RAPD分子标记;PCR扩增条件;DNA提取方法

外文关键词：Alnus cremastogyne; RAPD molecular marker; PCR

分类号：S792.14

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: predenaturating 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, dNTP 2.5 μL, primer (s60+s155) 2 mmol·L-1, Mg2+ 3.0 mmol·L-1, Tap enzyme 1 U, DNA 40 mg, then adding ddH2O 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: predenaturating 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, dNTP 2.5 μL, primer (s60+s155) 2 mmol·L-1, Mg2+ 3.0 mmol·L-1, Tap enzyme 1 U, DNA 40 mg, then adding ddH2O to 20 μL. This study may provide some references for the application of RAPD technique in genetic research of alder tree species.