期刊论文

A New Sampling Approach to Obtain Genomic DNA of Marine Shellfish

中文

安徽农业科学

0517-6611

2010

38

35

20112-20114

10.13989/j.cnki.0517-6611.2010.35.164

广西海洋生物技术重点实验室主任基金课题（GKLMBT-D0801）；

本校为第一机构

[目的]探讨一种新的获取海洋贝类基因组DNA的取样方法,为开展珍稀贝类的分子生物学研究提供参考。[方法]以文蛤、栉江瑶、翡翠贻贝、香港巨牡蛎、毛蚶为供试材料,以闭壳（合）肌全基因组DNA作为参考,采用常规酚-氯仿法抽提贝类贝腔液基因组DNA,使用生物光度计法、琼脂糖凝胶电泳、目的片段扩增测序验证其质量,并建立系统发育树检测其来源的真实可靠性。[结果]采用常规酚/氯仿基因组DNA提取方法获得的贝腔液DNA质量优于闭壳（合）肌DNA,蛋白质、多酚色素污染较少,完全满足目的片段扩增测序;系统发育进化树则证实了贝腔液并非外源污染。[结论]从贝腔液中获取基因组DNA是完全可行的,并可将其用于目...

[Objective] The aim was to explore a new sampling approach to obtain genomic DNA of marine shellfish,as well as to provide reference for the molecular biology research on precious shellfish.[Method] Meretrix meretrix,Atrina pectinata,Perna viridis,Crassostrea hongkongensis and Scapharca kagoshimensis were used as experimental materials and the genomic DNA of adductor muscle was taken as reference to extract the genomic DNA of shell cavity fluids with the conventional phenol-chloroform method.And then biophotometer,agarose gel electrophoresis,amplification and sequencing of the target fragments...