[1]张伟,谢甫绨,曹萍,等.大豆叶片DNA提取方法的比较研究[J].大豆科学,2007,26(01):60-65.[doi:10.3969/j.issn.1000-9841.2007.01.014]
　ZHANG Wei,XIE Fu-ti,CAO Ping,et al.COMPARATIVE STUDIES ON EXTRACTION OF GENOME DNA FROM SOYBEAN LEAF[J].Soybean Science,2007,26(01):60-65.[doi:10.3969/j.issn.1000-9841.2007.01.014]
点击复制

大豆叶片DNA提取方法的比较研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第26卷 期数: 2007年01期 页码: 60-65 栏目: 出版日期: 2007-02-25
Title:
COMPARATIVE STUDIES ON EXTRACTION OF GENOME DNA FROM SOYBEAN LEAF
文章编号:
1000-9841(2007)01-0060-06
作者:
张伟; 谢甫绨; 曹萍; 郭玉华; 宋显军
沈阳农业大学农学院，沈阳110161
Author(s):
ZHANG Wei; XIE Fu-ti; CAO Ping; GUO Yu-hua; SONG Xian-jun
College of Agronomy， Shenyang Agricultural University， Shenyang 110161
关键词:
DNA提取; SSR; CTAB; SDS
Keywords:
DNA extraction;  SSR;  CTAB;  SDS
分类号:
S565.103
DOI:
10.3969/j.issn.1000-9841.2007.01.014
文献标志码:
A
摘要:
以大豆叶片为材料，对大豆基因组DNA提取方法中若干影响因子如提取液、提取液的浓度、蛋白质去除次数、提取步骤等进行比较研究，试图寻找大豆叶片DNA提取的最佳方法。结果表明，碱裂解法提取速度快，但提取的DNA量少，在SSR分析中，银染效果较CTAB、SDS提取液差，CTAB、SDS提取液均能得到较高质量的DNA；在1%~4%CTAB、SDS提取液浓度下，4%CTAB易使DNA产生降解，但不同浓度提取液提取DNA均可用于SSR分析；提取叶片重量在一定范围内时，随抽提次数减少，DNA 浓度增加，纯度下降，但即使用氯仿/异戊醇抽提一次，也能够满足SSR分析的需要；不同提取步骤下，使用氯仿/异戊醇抽提2次后使DNA沉出后再溶解抽提2次和直接抽体4次相比，A260/A280值偏大，但A260/A230值较好。
Abstract:
Template DNA quality directly affects the effectiveness of PCR amplification. To investigate the optimum DNA extraction method from soybean leaf， the influencing factors such as extraction buffer， buffer concentration， extraction times， and extraction steps were analyzed. The results indicated that the alkali extraction method on leaf was faster than but inferior to the other two methods. Both CTAB and SDS method could produce high quality DNA. When the DNA quality was compared under different buffer concentration ranging from 1% to 4%， it was found the quality of DNA could meet need of SSR analysis except that the DNA degraded a little under 4% CTAB. The concentration of DNA increased as the extraction times decreased， while the purity of DNA descended. The quality of DNA could meet SSR requirement even extracted once by chloroform/isoamyl. Under different extraction steps， we extracted DNA 4 times continuously， or separated out DNA after extracting twice， then dissolved it and extracted twice more， compared to the former， the later had a bigger ratio of A260/A280， but a suitable ratio of A260/A230.

参考文献/References:

［1］周思君.小样品大批量大豆模板DNA快速分离［J］.大豆科学，1999，18（4）：318-321.

［2］王灏， 王道杰， 谭小力，等.用于RAPD分析的油菜总DNA 的快速提取［J］.西北农业学报，2001，10(3)：32- 34.
［3］戴思兰，陈俊愉，高荣孚，等.DNA提纯方法对9种菊属植物RAPD的影响［J］.园艺学报，1996，23 （2）：169-313.
［4］杜何为，黄敏，张祖新.玉米DNA的小量快速提取［J］.玉米科学，2004，12（2）：114-115.
［5］张永明，孙彩云，梁承邺.一步法提取植物DNA 用于大规模RAPD分析［J］.遗传，2000，22(2)：106.
［6］郭景伦，赵久然，辛景树，等.玉米单株幼芽DNA快速提取新方法［J］.华北农学报，2005，20（1）：38-40.
［7］Wang Shu，R E Knox，R M DePauw，J et al.A Simple DNA Preparation Method for PCR Amplifications in Marker.Assisted Selection of Wheat［J］.Agricultural Sciences in China，2005，4(7)：481-485.
［8］王晓丹，吕慧颖，张敬，等.以PCR为目的的大豆叶片DNA提取方法的比较研究［J］.分子植物育种，2004 2（6）：891-894.
［9］汪小全，皱育苹，张大明，等.RAPD应用于遗传多样性和系统学研究中的问题［J］.植物学报，1996，38（12）：954-962.
［10］卢阳江，郑康乐.提取水稻DNA的一种简易方法［J］.中国水稻科学，1992，6（1）：47-48.
［11］郑康乐，钱慧荣，庄杰云，等.应用DNA标记定位水稻抗稻瘟病基因［J］.植物病理学报，1995，25（4）：307-313.

相似文献/References:

[1]吴 禹,沈 军,陈爱国,等.辽宁省野生大豆资源遗传多样性的比较分析[J].大豆科学,2012,31(03):368.[doi:10.3969/j.issn.1000-9841.2012.03.007]
　WU Yu,SHEN Jun,CHEN Ai-guo,et al.Genetic Similarity for Wild Soybeans from Different Geographical Origins in Liaoning Province[J].Soybean Science,2012,31(01):368.[doi:10.3969/j.issn.1000-9841.2012.03.007]
[2]曾维英,梁江,陈渊,等.广西新收集野生大豆资源的遗传多样性分析[J].大豆科学,2011,30(03):379.[doi:10.11861/j.issn.1000-9841.2011.03.0379]
　ZENG Wei-ying,LIANG Jiang,CHEN Yuan,et al.Genetic Diversity Analysis of New Wild Soybean Collection in Guangxi[J].Soybean Science,2011,30(01):379.[doi:10.11861/j.issn.1000-9841.2011.03.0379]
[3]李建东,燕雪飞,董思言,等.辽宁省野生大豆种质资源的SSR遗传多样性分析[J].大豆科学,2010,29(01):28.[doi:10.11861/j.issn.1000-9841.2010.01.0028]
　LI Jian-dong,YAN Xue-fei,DONG Si-yan,et al.Analysis of Genetic Diversity of Glycine soja Germplasm Resources in Liaoning Province[J].Soybean Science,2010,29(01):28.[doi:10.11861/j.issn.1000-9841.2010.01.0028]
[4]盛新颖,詹少华,樊洪泓,等.利用SSR分子标记鉴定“梅桥”大豆种质纯度[J].大豆科学,2010,29(02):210.[doi:10.11861/j.issn.1000-9841.2010.02.0210]
　SHENG Xin-ying,ZHAN Shao-hua,FAN Hong-hong,et al.Purity Identification of “Meiqiao” Soybean (Glycine max) by SSR Markers[J].Soybean Science,2010,29(01):210.[doi:10.11861/j.issn.1000-9841.2010.02.0210]
[5]王振东,陈超力,于佰双,等.大豆抗旱种质资源遗传多样性的SSR分析[J].大豆科学,2010,29(03):370.[doi:10.11861/j.issn.1000-9841.2010.03.0370]
　WANG Zhen-dong,CHEN Chao-li,YU Bai-shuang,et al.SSR Analysis of Genetic Diversity of Soybean Germplasm in Drought Resistance[J].Soybean Science,2010,29(01):370.[doi:10.11861/j.issn.1000-9841.2010.03.0370]
[6]王丹,乔亚科,韩粉霞,等.河北东部沿海地区野生大豆SSR多样性分析[J].大豆科学,2010,29(04):555.[doi:10.11861/j.issn.1000-9841.2010.04.0555]
　WANG Dan,QIAO Ya-ke,HAN Fen-xia,et al.Genetic Diversity of Glycine soja in Eastern Coastal Area of Hebei Province[J].Soybean Science,2010,29(01):555.[doi:10.11861/j.issn.1000-9841.2010.04.0555]
[7]周斌,邢邯,陈受宜,等.大豆分子标记遗传图谱的整合及其应用[J].大豆科学,2009,28(04):557.[doi:10.11861/j.issn.1000-9841.2009.04.0557]
　ZHOU Bin,XING Han,CHEN Shou-yi,et al.An Integrated Genetic Linkage Map of Soybean and Its Application[J].Soybean Science,2009,28(01):557.[doi:10.11861/j.issn.1000-9841.2009.04.0557]
[8]陈远东,喻德跃.EMS诱发大豆“南农94-16”突变体库的扩建及部分突变体的SSR分析[J].大豆科学,2009,28(04):574.[doi:10.11861/j.issn.1000-9841.2009.04.0574]
　CHEN Yuan-dong,YU De-yue.Construction of Mutant Pools for Soybean “Nannong 94-16” Induced by EMS and Analysis of SSR Marker on Several Mutants[J].Soybean Science,2009,28(01):574.[doi:10.11861/j.issn.1000-9841.2009.04.0574]
[9]陈相艳,李伟,戴海英,等.大豆EST资源的SSR信息分析[J].大豆科学,2009,28(03):394.[doi:10.11861/j.issn.1000-9841.2009.03.0394]
　CHEN Xiang-yan,LI Wei,DAI Hai-ying,et al.Analysis of SSR Information in EST Resource of Soybean（Glycine max)[J].Soybean Science,2009,28(01):394.[doi:10.11861/j.issn.1000-9841.2009.03.0394]
[10]王俊,王林林,刘章雄,等.大豆地方品种遗传结构及其保存研究[J].大豆科学,2008,27(03):361.[doi:10.11861/j.issn.1000-9841.2008.03.0361]
　WANG Jun,WANG Lin-lin,LIU Zhang-xiong,et al.Genetic Structure and Conservation of Soybean Landraces[J].Soybean Science,2008,27(01):361.[doi:10.11861/j.issn.1000-9841.2008.03.0361]

备注/Memo

基金项目：国家948项目“大豆优良种质资源和促进先进技术的引进与创新”资助（2003-Q04)

作者简介：张伟（1979-），男，在读博士，从事大豆产量生理研究。

更新日期/Last Update: 2014-10-21