一种使用十六烷基三甲基溴化铵从植物组织中快速提取DNA的改良方法。

A modified protocol for rapid DNA isolation from plant tissues using cetyltrimethylammonium bromide.

作者信息

Allen G C, Flores-Vergara M A, Krasynanski S, Kumar S, Thompson W F

机构信息

Department of Horticultural Science and Crop Science, 1200 Partners II, Campus Box 7550, 840 Main Campus Drive, North Carolina State University, Raleigh, NC 27606-7550, USA.

出版信息

Nat Protoc. 2006;1(5):2320-5. doi: 10.1038/nprot.2006.384.

DOI:10.1038/nprot.2006.384

PMID:17406474

Abstract

We describe a modification of the DNA extraction method, in which cetyltrimethylammonium bromide (CTAB) is used to extract nucleic acids from plant tissues. In contrast to the original method, the modified CTAB procedure is faster, omits the selective precipitation and CsCl gradient steps, uses less expensive and toxic reagents, requires only inexpensive laboratory equipment and is more readily adapted to high-throughput DNA extraction. This protocol yields approximately 5-30 microg of total DNA from 200 mg of tissue fresh weight, depending on plant species and tissue source. It can be completed in as little as 5-6 h.

摘要

我们描述了一种DNA提取方法的改进，其中使用十六烷基三甲基溴化铵（CTAB）从植物组织中提取核酸。与原始方法相比，改进后的CTAB方法更快，省略了选择性沉淀和CsCl梯度步骤，使用的试剂成本更低且毒性更小，只需要价格便宜的实验室设备，并且更易于适应高通量DNA提取。根据植物种类和组织来源，该方案从200毫克组织鲜重中可产生约5 - 30微克的总DNA。整个过程最短可在5 - 6小时内完成。