HUANGWen-gong.Optimizing the RAPD Reaction System of Flax and Screening Primers[J].HEILONGJIANG AGRICULTURAL SCIENCES,2009,(02):4-6.
亚麻RAPD 的反应体系优化及引物筛选
- Title:
- Optimizing the RAPD Reaction System of Flax and Screening Primers
- 文章编号:
- 1002-2767(2009)02-0004-03
- Keywords:
- flax ; genome DNA; RAPD reaction ; screen
- 分类号:
- S563.2
- 文献标志码:
- A
- 摘要:
- 对亚麻RAPD 反应条件进行了优化:在25 μL 反应体系中, 含10×Buffer , Mg2+(1 mmol·L-1), Taq 酶1 U, GenomeDNA 50 ng , dNTP 0.25 mmol·L-1 , RAPD primer 1.5 μmol·L-1 。PCR 反应程序为:94℃预变性4 min;94℃变性40s, 37℃退火1 min ,72 ℃延伸90 s, 40 个循环;72℃延伸10 min。同时利用3 个有代表性的亚麻品种从70 条引物中筛选出12 条多态性好、重复性高的引物, 为亚麻的分子鉴定奠定了基础。
- Abstract:
- This experiment o ptimized the RAPD reaction conditions of flax .During 25 μL reaction system, it contained 10×Buffer , Mg2 +(1 mmol·L-1), Taq enzyme 1 U, Genome DNA 50 ng , dNTP 0.25 mmol·L-1 and RAPD primer 1.5 μmol·L-1 .The process of PCR were 94℃ predenatalizating for 4 min, 40 cycles(94℃ denatalizating for 40 s, 37 ℃ annealing for 1min, 72℃ extending for 90 s), 72℃ ex tending fo r 10 min.Meanwhile, the experiment used 3 typical flax varieties to screen12 primers which had high level polymorphism and repeatability from 70 primers.This research made the foundation for themolecular identification of flax.
参考文献/References:
[ 1] Wil li am s J G .DNA polymorphisms ampli fied by arbit raryprimers are useful as geneti c markers[ J] .Nucl eic AcidsRes ,1990 , 18 :6531-6535 .[ 2] Wel sh J , M cClelland M .Gen om e f ingerprint ing u sing PCR w itharbi t rary primers[ J] .Nu clei c Acids Res , 1990 , 8 :7213-7218 .[ 3] 王关林, 方宏筠.植物基因工程原理与技术[ M] .北京:科学出版社, 1998:598-599 .[ 4] 萨姆布鲁克J , 拉塞尔D W.分子克隆实验指南[ M] .北京:科学出版社,2002 .
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备注/Memo
收稿日期:2008-05-27基金项目:国家“十一五” 科技支撑计划项目;黑龙江省农业科技创新工程青年基金项目作者简介:黄文功(1980-), 男, 黑龙江省哈尔滨市人, 硕士,研究实习员, 主要从事亚麻分子育种方面研究。Tel :0451-86677430;E-mail:huangwengong1736 @yahoo.com .cn