从植物中提取古代DNA。

Ancient DNA extraction from plants.

作者信息

Kistler Logan

机构信息

Department of Anthropology, The Pennsylvania State University, 409 Carpenter Building, University Park, PA 16802, USA.

出版信息

Methods Mol Biol. 2012;840:71-9. doi: 10.1007/978-1-61779-516-9_10.

DOI:10.1007/978-1-61779-516-9_10

PMID:22237524

Abstract

A variety of protocols for DNA extraction from archaeological and paleobotanical plant specimens have been proposed. This is not surprising given the range of taxa and tissue types that may be preserved and the variety of conditions in which that preservation may take place. Commercially available DNA extraction kits can be used to recover ancient plant DNA, but modifications to standard approaches are often necessary to improve yield. In this chapter, I describe two protocols for extracting DNA from small amounts of ancient plant tissue. The CTAB protocol, which I recommend for use with single seeds, utilizes an incubation period in extraction buffer and subsequent chloroform extraction followed by DNA purification and suspension. The PTB protocol, which I recommend for use with gourd rind and similar tissues, utilizes an overnight incubation of pulverized tissue in extraction buffer, removal of the tissue by centrifugation, and DNA extraction from the buffer using commercial plant DNA extraction kits.

摘要

已经提出了多种从考古和古植物学植物标本中提取DNA的方案。鉴于可能保存的分类群和组织类型的范围以及保存可能发生的各种条件，这并不奇怪。市售的DNA提取试剂盒可用于回收古代植物DNA，但通常需要对标准方法进行修改以提高产量。在本章中，我描述了两种从少量古代植物组织中提取DNA的方案。我推荐用于单粒种子的CTAB方案，它在提取缓冲液中孵育一段时间，随后进行氯仿提取，然后进行DNA纯化和悬浮。我推荐用于瓜皮和类似组织的PTB方案，它将粉碎的组织在提取缓冲液中孵育过夜，通过离心去除组织，并使用商业植物DNA提取试剂盒从缓冲液中提取DNA。