摘要
对影响水稻SRAP-PCR扩增结果的模板质量浓度、dNTP Mixture浓度、Mg2+浓度以及引物、反应程序进行了探索,获得在使用大连宝生物Taq酶时能够稳定扩增水稻基因组的SRAP-PCR体系:在25μL体系中,合适的模板DNA质量浓度为0.8~1.2 ng/μL,Mg2+浓度为2~3 mmol/L,dNTP浓度为0.15~0.20mmol/L,按照Li和Quiros设计的程序或按照"94℃预变性5 min;94℃变性1 min,35℃复性1 min,72℃延伸1min,35循环,最后72℃延伸10 min,4℃保存"的程序进行扩增可以得到较为满意的结果.Li和Quiros设计的30对引物均能用于水稻SRAP-PCR.
Some factors involving concentration of template,concentration of dNTP mixture,concentration of Mg2+ and primer,reactive program of PCR of rice were studied and an advanced technical system was established.The 25 μL reaction mixture contained:0.8—1.2 ng/μL of genomic DNA template,2—3 mmol/L of Mg2+,0.15—0.20 mmol/L of dNTP and appropriate reaction buffer.Two reactive programs can be used,which is the program of design by Li and Quiros or "initially denaturing at 94 ℃ for 5 min;then denaturation at 94 ℃,1 mi...
出处
《云南大学学报（自然科学版）》
CAS
CSCD
北大核心
2008年第S1期421-425,共5页
Journal of Yunnan University(Natural Sciences Edition)
基金
海南省自然科学研究基金资助项目(30514)
