HAN Wei-bo,TANG Feng-lan,ZHANG Yue-xue,et al.Optimalization of RAPD Reaction System for Clover[J].HEILONGJIANG AGRICULTURAL SCIENCES,2006,(01):10-12.[doi:10.11942/j.issn1002-2767.2006.01.0010]
三叶草RAPD 反应条件的优选
- Title:
- Optimalization of RAPD Reaction System for Clover
- 文章编号:
- 1002 - 2767(2006)01 - 0010 - 03
- Keywords:
- clover ; RAPD ; amplified reaction ; optimalization
- 分类号:
- S541. 2
- 文献标志码:
- A
- 摘要:
- 以红三叶( Tri f ol ium pretense L. )、白三叶(Tri f ol ium repens L . ) 和杂三叶(Tri f ol iumHybrid um L. )三种三叶草为材料, 对RAPD 反应条件中的dN TP 浓度、随机引物浓度、模板DNA用量、Taq DNA 聚合酶用量、退火温度以及预变性时间分别进行梯度试验。结果表明, 该反应体系体积为25 μL :ddH2O , 19. 1 μL ;10 ×Buf fer , 2. 5 μL ;Primer , 200 nM ;dNTP , 200 μM ;Taq DN A 聚合酶, 1U ;Template DNA 1ng /μL 。通过试验建立了一套稳定的RAPD 反应体系, 该反应体系扩增效果良好。
- Abstract:
- Three specie s of clover w ere used as the material s o f ex t ract ing DNA in this experiment, they w ere T rifolium pretense L. 、T rifolium repens L. and Trifolium Hybridum L. . Thee xperiment step to the concent rat ion of dN TP 、random primers 、template DNA 、Taq DNA polymerase、annealing temperature and preheat denaturalizatio n time on RA PD w ere conducted. Thena stable reactio n sy stem of RAPD w as set up. In the optimalization of RAPD sy stem wi th to talevolume o f 25μL :ddH2O , 19. 1μL ;10 ×Buf fer , 2. 5μL ;Primer , 200nM ;dN TP , 200μM ;Taq DNAE , 1U ;Template DNA 1ng /μL. The effect of amplifying w ith the reaction sy stem w as the mo stoptimal.
参考文献/References:
[ 1]JIANG Yan - xi , ZH ANG Yue - xue, TANG Feng - lan , etal. . The Gen et ics Study of Alf al fa Irradi at ed in Magneti c FieldFree S pace[ J]. china bi otechn ology ,2005 , 25(增):148-150.
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备注/Memo
收稿日期:2005 - 10 - 19