新疆甜瓜AFLP反应体系的建立

Establishment of AFLP Technical System in Xinjiang Melon

以新疆甜瓜叶片为试验材料,通过对扩增片段长度多态性(AFLP)反应体系中的几个关键参数进行研究,建立了适合新疆甜瓜的AFLP技术体系.结果表明,采用改进CTAB法提取的基因组DNA质量好,符合AFLP反应要求;双酶切必须完全彻底,基因组DNA 300 ng用EcoR I和MseI各3 U,37℃酶切5 h;连接产物稀释10倍用于预扩增;预扩增产物的稀释以75倍为宜.本研究为构建新疆甜瓜的遗传图谱及分子标记辅助育种奠定了基础.

An AFLP （amplified fragment length polymorphism） analysis system was established with Xinjiang melon leaves by optimizing some factors of the AFLP technical system. The results showed that the modified CTAB could be used for extracting ideal DNA and be suitable for AFLP, the double digestion should be complete,300 ng genomic DNA was digested with 3 U ofEcoR I and 3 U of MseI at 37℃ for 5 hours. The digestedligated DNA fragment was diluted （10 folds） and used as template for pre-amplification, and the products of pre-ampllflcation should be diluted to 75 times. This study has laid a foundation for the construction of genetic map and molecular marker-assisted breeding in Xinjiang melon.