广西植物
廣西植物
엄서식물
GUIHAIA
2010年
1期
112-116
,共5页
胡延萍%谢小龙%王莉%杨建%李毅
鬍延萍%謝小龍%王莉%楊建%李毅
호연평%사소룡%왕리%양건%리의
唐古特大黄%ISSR-PCR%优化
唐古特大黃%ISSR-PCR%優化
당고특대황%ISSR-PCR%우화
Rheum tanguticum%ISSR-PCR%optimization
利用单因素试验对影响唐古特大黄ISSR-PCR扩增的重要参数进行优化,以期建立其最佳反应条件.结果如下:20 μL反应体系包括1.5×PCR buffer(15 mmol/L Tris-HCl,75 mmol/L KCl),1.00 mmol/L MgCl_2,0.6U Taq DNA聚合酶,0.125 mmol/L dNTP,0.5 μmol/L引物和30 ng模板DNA;引物UBC888适宜的退火温度为57.4 ℃.ISSR反应条件的建立为利用分子标记技术研究唐古特大黄居群遗传多样性奠定了良好基础.
利用單因素試驗對影響唐古特大黃ISSR-PCR擴增的重要參數進行優化,以期建立其最佳反應條件.結果如下:20 μL反應體繫包括1.5×PCR buffer(15 mmol/L Tris-HCl,75 mmol/L KCl),1.00 mmol/L MgCl_2,0.6U Taq DNA聚閤酶,0.125 mmol/L dNTP,0.5 μmol/L引物和30 ng模闆DNA;引物UBC888適宜的退火溫度為57.4 ℃.ISSR反應條件的建立為利用分子標記技術研究唐古特大黃居群遺傳多樣性奠定瞭良好基礎.
이용단인소시험대영향당고특대황ISSR-PCR확증적중요삼수진행우화,이기건립기최가반응조건.결과여하:20 μL반응체계포괄1.5×PCR buffer(15 mmol/L Tris-HCl,75 mmol/L KCl),1.00 mmol/L MgCl_2,0.6U Taq DNA취합매,0.125 mmol/L dNTP,0.5 μmol/L인물화30 ng모판DNA;인물UBC888괄의적퇴화온도위57.4 ℃.ISSR반응조건적건립위이용분자표기기술연구당고특대황거군유전다양성전정료량호기출.
Inter-Simple Sequence Repeats(ISSR)is a good molecular marker for revealing genetic diversity. Reaction system differed in different species,so optimization of ISSR-PCR reaction is very important. Factors which affect the ISSR-PCR amplification,such as the concentration of Mg~(2+),Taq DNA polymerase,dNTP,primer and template DNA with different annealing temperatures,were optimized and selected by using the genomic DNA of Rheum tanguticum as material. Optimal PCR(20 μL)mix contained 1.5×PCR buffer(15 mmol/L Tris-HCl,75 mmol/L KCl),1.00 mmol/L MgCl_2,0.6 U Taq DNA polymerase,0.125 mmol/L dNTP,0.5 μmol/L primer and 30ng template DNA. The suitable annealing temperature was 57.4 ℃ for primer UBC888. Establishment of the PCR reaction conditions could favor further studies on the genetic diversity of R.tanguticum by using ISSR molecular marker techniques.
