高良姜AFLP技术体系的建立与优化

Establishment and Optimization of AFLP Technical System of Alpinia officinarum Hance

为了建立适合高良姜的扩增片段长度多态性(AFLP)技术体系,对AFLP技术中主要环节的反应条件进行了优化。研究结果表明,用改良CTAB法可提取高质量的高良姜基因组DNA,基因组DNA用限制性内切酶EcoRⅠ和MseⅠ各4U于37℃水浴2h可以被完全酶切,酶切片段与EcoRⅠ和MseⅠ接头在16℃连接过夜后,用优化的反应体系进行预扩增和选择性扩增,扩增产物经过6%变性聚丙烯酰胺凝胶电泳和银染后获得清晰的多态性指纹图谱,因此说明所建立的AFLP技术体系符合要求。

In order to establish a suitable AFLP technical system for A. officinarum Hance, the reaction conditions of the main steps of AFLP technology were optimized. The results indicated that high quality genomic DNA was extracted by the modified CTAB method, and the genomic DNA was completely digested by 4U enzyme EcoR I and Mse I , respectively, in 37 ℃ water-bath for 2 h. The digested fragments were legated with EcoR l and Mse I adaptors at 16℃ to stand overnight, and then were performed through pre-amplification and selective amplification in optimized reaction conditions. The amplification products were analyzed in 6% denaturing polyacrylamide gel. Limpid polymorphism fingerprinting was obtained after silver staining, so, the AFLP technical system satisfied the demand.