[1]胡薇,陈宇,林来水,等.黑木相思基因组DNA的快速提取[J].华侨大学学报(自然科学版),2009,30(1):71-74.[doi:10.11830/ISSN.1000-5013.2009.01.0071]
　HU Wei,CHEN Yu,LIN Lai-shui,et al.Fast Extraction of Genome DNA from Acacia melanoxylon[J].Journal of Huaqiao University(Natural Science),2009,30(1):71-74.[doi:10.11830/ISSN.1000-5013.2009.01.0071]
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黑木相思基因组DNA的快速提取()
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《华侨大学学报（自然科学版）》[ISSN:1000-5013/CN:35-1079/N]

卷:
第30卷
期数:
2009年第1期
页码:
71-74
栏目:
出版日期:
2009-01-20

文章信息/Info

Title:
Fast Extraction of Genome DNA from Acacia melanoxylon
文章编号:
1000-5013(2009)01-0071-04
作者:
胡薇; 陈宇; 林来水; 林思祖
福建农林大学林学院
Author(s):
HU Wei;  CHEN Yu;  LIN Lai-shui;  LIN Si-zu
College of Forestry, Fujian Agriculture and Forestmg University, Fuzhou 350002, China
关键词:
黑木相思;  基因组DNA;  提取;  CTAB法
Keywords:
Acacia melanoxylon;  genomic;  DNA;  extraction;  CTAB method
分类号:
S792.99; Q943.2
DOI:
10.11830/ISSN.1000-5013.2009.01.0071
文献标志码:
A
摘要:
用SDS高盐介质法和1.5×CTAB法提取黑木相思幼叶及叶状柄基因组DNA,将其与2×CTAB法、3×CTAB法、改进的CTAB法进行比较,并用紫外光谱吸收、凝胶电泳、限制性内切酶消化、RAPD(Radomly Amplified Polymorphic DNA)和ISSR(Inter-Simple Sequence Repeat)等方法进行鉴定.结果表明,叶状柄多糖的质量分数较幼叶少,更易于基因组DNA的提取; SDS高盐介质法与1.5×CTAB法提取叶状柄的基因组DNA得率为180.0~697.5μg.g-1,分子质量为48 kb,D(260)/D(280)=1.750~1.955,无需经RNase处理,可直接用于限制性酶切和RAPD及ISSR分析; 1.5×CTAB法所提的基因组DNA的纯度优于SDS高盐介质法.综合考虑认为,1.5×CTAB法可作为黑木相思基因组DNA快速提取的简便方法.
Abstract:
A fast and convenient method for extraction of DNA from normal young fresh leaves and phyllode of Acacia melanoxylon was introduced by comparing SDS high salt method and 1.5×CTAB method with 2×CTAB,3×CTAB and ameliorated CTAB method,and tests were carried out with ultraviolet spectrophotometer analysis,agarose gel electrophoresis,restriction enzyme digestion,RAPD and ISSR reaction respectively.The results indicated that 180~697.5 μg·g-1 of DNA was obtained from a phyllode,their molecular weights were about 48 kb,and the D(260)/D(280) of DNA was 1.750~1.955,the DNA was suitable for digesting with restriction enzyme,RAPD and ISSR reaction without purifying by RNase,the amylose in phyllode is much less than that in normal young fresh leaves,and extraction of genome DNA from phyllode is easier than normal young fresh leaves,the quality of DNA obtained with 1.5×CTAB method is better than SDS high salt method,It is suggested that 1.5×CTAB method can be considered as one of fast and convenient method for extraction of DNA in young fresh leaves of Acacia melanoxylon.

参考文献/References:

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备注/Memo

备注/Memo:
福建省自然科学基金资助项目(B0410007)
更新日期/Last Update: 2014-03-23