从淹没的松木中提取DNA。

Extracting DNA from submerged pine wood.

作者信息

Reynolds M Megan, Williams Claire G

机构信息

Graduate Genetics Program, Texas A&M University, College Station, TX 77844, USA.

出版信息

Genome. 2004 Oct;47(5):994-7. doi: 10.1139/g04-045.

DOI:10.1139/g04-045

PMID:15499414

Abstract

A DNA extraction protocol for submerged pine logs was developed with the following properties: (i) high molecular weight DNA, (ii) PCR amplification of chloroplast and nuclear sequences, and (iii) high sequence homology to voucher pine specimens. The DNA extraction protocol was modified from a cetyltrimehtylammonium bromide (CTAB) protocol by adding stringent electrophoretic purification, proteinase K, RNAse, polyvinyl pyrrolidone (PVP), and Gene Releaser. Chloroplast rbcL (ribulose-1,5-bisphosphate carboxylase) could be amplified. Nuclear ribosomal sequences had >95% homology to Pinus taeda and Pinus palustris. Microsatellite polymorphism for PtTX2082 matched 2 of 14 known P. taeda alleles. Our results show DNA analysis for submerged conifer wood is feasible.

摘要

已开发出一种用于水下松木原木的DNA提取方案，其具有以下特性：（i）高分子量DNA，（ii）叶绿体和核序列的PCR扩增，以及（iii）与凭证松标本的高序列同源性。通过添加严格的电泳纯化、蛋白酶K、RNA酶、聚乙烯吡咯烷酮（PVP）和基因释放剂，对十六烷基三甲基溴化铵（CTAB）方案进行了修改，从而得到该DNA提取方案。叶绿体rbcL（核酮糖-1,5-二磷酸羧化酶）能够被扩增。核糖体核序列与火炬松和湿地松的同源性大于95%。PtTX2082的微卫星多态性与14个已知火炬松等位基因中的2个相匹配。我们的结果表明，对水下针叶树木材进行DNA分析是可行的。