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中国樱桃S-RNase基因PCR扩增技术体系的建立
引用本文:李晓,吴俊,张绍铃.中国樱桃S-RNase基因PCR扩增技术体系的建立[J].果树学报,2008,25(2):277-280.
作者姓名:李晓  吴俊  张绍铃
作者单位:南京农业大学园艺学院,南京,210095
摘    要:通过对TaqDNA聚合酶、Mg2+、dNTP、通用引物、模板DNA浓度等反应参数的系统研究,建立了中国樱桃S-RNase基因特异PCR扩增体系。该体系反应的总体积25μL,其中Taq酶1.25U,MgCl22.5mmol/L,dNTP0.15mmol/L,通用引物0.25μmol/L,模板DNA 50 ng。反应程序为:94℃预变性3 min;33个循环的94℃变性1 min,56℃退火45 s,72℃延伸1.5 min;最后72℃延伸10 min。利用优化的扩增体系在中国樱桃的不同品种中获得有效扩增,进一步证明了该体系具有良好的稳定性和重复性。

关 键 词:中国樱桃  S-RNase基因  PCR扩增体系
文章编号:1009-9980(2008)02-277-04
收稿时间:2007-07-21
修稿时间:2008-01-11

Establishment of PCR reaction system of S-RNase gene in Chinese cherry cultivars ( Cerasus pseudocerasus )
LI Xiao,WU Jun,ZHANG Shao-ling.Establishment of PCR reaction system of S-RNase gene in Chinese cherry cultivars ( Cerasus pseudocerasus )[J].Journal of Fruit Science,2008,25(2):277-280.
Authors:LI Xiao  WU Jun  ZHANG Shao-ling
Abstract:A special PCR reaction system has been established for S-RNase gene in Chinese cherry through the optimization of the concentration of Taq DNA polymerase,Mg2+,dNTP,universal primers and template DNA.The reaction volume was 25 μL,containing 1.25 U Taq DNA polymerase,2.5 mmol/L MgCl2,0.15 mmol/L dNTP,0.25 μmol/L universal primers and 50 ng DNA.The PCR cycle was designed as following,pre-denaturing at 94 ℃ for 3 min,denaturing at 94 ℃ for 1 min,annealing at 56 ℃ for 45 s,extension at 72 ℃ for 1.5 min,total 33 cycles,then extension at 72 ℃ for 10 min.The stability and repeatability of the system had been validated by the effective amplification of S-RNase genes in Chinese cherry cultivars.
Keywords:Chinese cherry  S-RNase gene  PCR amplification system
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