棉 花 学 报    Cotton Science   2010，22（3）：202-208

The Optimization of ISSR-PCR System of Cotton by Uniform Design
ZHU Yan-fang¹，ZHU Shui-jin¹，WANG Yang¹，LI Yong-ping²，MA Wen-guang²，HU Jin^1*
(1.Seed Science Center of Agriculture and Biotechnology College，Zhejiang University，Hangzhou 310029，China； 2.Southern Center of China Tobacco Breeding Research，Yuxi, Yunnan 653100，China)

Abstract: In present study，ISSR-PCR reaction conditions on Gossypium spp. were optimized. The factors which affect ISSR results 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 μ mol·L^-1 primer and 2.0 mmol·L^-1 Mg²⁺ in 20 μ L 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.
Key words: cotton；ISSR；system optimization；uniform design    [Full Text, 468KB]