摘要
为探索适宜棉花的ISSR-PCR反应体系,采用两轮均匀设计试验,对ISSR-PCR反应体系中Mg2+、dNTPs、模板DNA、引物、Taq DNA聚合酶的浓度或用量进行优化。结果表明,棉花20μL的ISSR反应体系的最佳组分包括2μL 10×PCR Buffer、0.5 U Taq DNA聚合酶、40 ng模板DNA、0.25 mmol.L-1 dNTPs、0.5μmol.L-1引物和2.0 mmol.L-1 Mg2+。利用8个棉花品种和三条不同引物验证该反应体系,结果表明,该反应体系的稳定性和重复性较好。
In present study,ISSR-PCR reaction conditions on Gossypium spp.were optimized.The factors which affect ISSR re-sults of Gossypium spp.were investigated by uniform designs of two times.The results showed that 0.5 U Taq polymerase,40 ng template DNA,0.25 mmol.L-1 dNTPs,0.5 μmol.L-1 primer and 2.0 mmol.L-1 Mg2＋ in 20 μL ISSR reaction system might be the best combination.To validate the ISSR-PCR reaction system above,eight cultivars of Gossypium and three ISSR primers were employed in the research.It is indicated that the ISSR reaction system would be more productive and repeatable
出处
《棉花学报》
CSCD
北大核心
2010年第3期202-208,共7页
Cotton Science
基金
浙江省重大科技专项(优先主题)(2008C12005-1)
农业部转基因生物新品种培育重大专项(2008ZX08005-005)
云南省科技厅项目(2009EB073)
云南省烟草公司项目(09YN008)资助
