HAO Ruo-chao,QU Chun-l i,ZHANG Yue-xue,et al.Study on the Methods of Extraction and Purification for Alfalfa DNA and the Establishment and Optimization for RAPD Reaction System[J].HEILONGJIANG AGRICULTURAL SCIENCES,2006,(05):89-92.[doi:10.11942/j.issn1002-2767.2006.05.0089]
苜蓿基因组DNA 的提取及RAPD 反应组成优化探讨
- Title:
- Study on the Methods of Extraction and Purification for Alfalfa DNA and the Establishment and Optimization for RAPD Reaction System
- 文章编号:
- 1002 - 2767(2006)05 - 0089 - 04
- Keywords:
- alfalfa ; DNA ext raction ; RAPD react ion
- 分类号:
- S551. 7
- 文献标志码:
- A
- 摘要:
- 比较了CTAB 法和SDS 法提取苜蓿基因组总DNA 的效果, 这两种方法得到的全基因DNA具有较好的完整性, 但从DNA 的提取纯度可以看出, CTAB 法优于SDS 法。实验对苜蓿RAPD反应条件进行了优化:在25μL 反应体系中, 含10 ×Buf fer(20mM MgCl2 ), 1U Taq 酶, 25ng 的模板DNA , 150 mol /L 的dN TPs , 0. 2 mol /L 引物。PCR 反应程序为:94 ℃预变性3 min ;94 ℃变性15 s , 37 ℃退火30 s , 72 ℃延伸1 min , 35 个循环;72 ℃延伸10 min ;最后4 ℃保存。
- Abstract:
- Two me thods (SDS method and C TAB method) fo r DNA ex t racting w ere compared ,and w hole DNA co uld be g ot by tw o metho ds. But took purity of DNA into account , CTABmethod w as bet ter than SDS method. Meantime , PCR amplif ied procedure w as establi shed andPCR amplif ied co nditio ns w ere defined on the factors of Taq DNA polymerase , DNA densi ty ,dN TPs density , primer density , and pre-denaturating temperature.
参考文献/References:
[ 1]向育君, 梁辉, 张正. 稗草DNA 的快速提取研究[ J] . 生物学杂志, 2004 ,(2):31-34.
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备注/Memo
收稿日期:2006 - 04 - 05