单条线虫DNA提取方法研究

doi:10.7668/hbnxb.2014.02.024

华北农学报 ›› 2014, Vol. 29 ›› Issue (2): 127-132. doi: 10.7668/hbnxb.2014.02.024

单条线虫DNA提取方法研究

容万韬^1,2, 王金成², 刘鹏², 林宇², 郭京泽², 黄国明², 孙建华¹

1 天津师范大学生命科学学院天津市动植物抗性重点实验室天津 300387;
2 天津出入境检验检疫局动植物与食品检测中心天津 300461

收稿日期:2013-11-24 出版日期:2014-04-28
通讯作者: 孙建华(1955-),女,天津人,教授,博士,主要从事真菌代谢产物杀线虫活性物质的发酵、分离及防治植物线虫病害的研究。
作者简介:容万韬(1987-),男,广西钦州人,硕士,主要从事植物寄生线虫鉴定和生物防治研究。
基金资助:
国家自然科学基金项目(31272019);国家质检总局项目(2011IK176);国家科技支撑计划项目(2012BAK11B03)

Research on DNA Extraction Methods from a Single Nematode

RONG Wan-tao^1,2, WANG Jin-cheng², LIU Peng², LIN Yu², GUO Jing-ze², HUANG Guo-ming², SUN Jian-hua¹

1 College of Life Sciences, Tianjin Normal University, Tianjin Key Laboratory of Animals and Plants Resistance, Tianjin 300387, China;
2 Animal & Plant & Food Stuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau, Tianjin 300461, China

Received:2013-11-24 Published:2014-04-28

摘要/Abstract

摘要： 为探寻一种高效、稳定、廉价的单条线虫DNA提取方法,以6种植物寄生线虫作为试验材料,比较了4种方法,即直接裂解法、冻融裂解法、切割裂解法、切割冻融裂解法对单条线虫DNA的提取效果。结果表明,对线虫进行管内切割的切割冻融裂解法与切割裂解法高效、稳定,冻融裂解法效果较差,直接裂解法效果最差。对线虫进行管内切割的切割冻融裂解法和切割裂解法可高效、稳定地获得高质量的单条线虫总DNA,满足下一步的PCR扩增需求。

关键词: 蛋白酶K, 裂解, 冻融, 管内切虫, DNA提取

Abstract: To explore an efficient, stable, low-cost DNA extraction method from single nematode for the support of molecular identification of nematodes, using six species of plant-parasitic nematodes as experimental materials, four DNA extraction methods, namely, direct lysis method, freeze-thaw lysis method, cutting lysis method, freeze-thaw cutting lysis method were designed and compared in order to check their efficiency of DNA extraction from single nematode. The results showed that cutting lysis method and freeze-thaw cutting lysis method were the most efficient and stable methods for DNA extraction of single nematode, then the freeze-thaw lysis method, and the direct lysis method was the worst one. Total DNA with high-quality from a single nematode could be gained efficiently and stab- ly through two methods, namely, cutting lysis method and freeze-thaw cutting lysis method. And the extracted DNA meeted requirement of the following PCR amplification.

Key words: Proteinase K, Lysis, Freeze-thaw, Nematode-cutting in tube, DNA extraction

中图分类号:

S154.38+6

容万韬, 王金成, 刘鹏, 林宇, 郭京泽, 黄国明, 孙建华. 单条线虫DNA提取方法研究[J]. 华北农学报, 2014, 29(2): 127-132. doi: 10.7668/hbnxb.2014.02.024.

RONG Wan-tao, WANG Jin-cheng, LIU Peng, LIN Yu, GUO Jing-ze, HUANG Guo-ming, SUN Jian-hua. Research on DNA Extraction Methods from a Single Nematode[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(2): 127-132. doi: 10.7668/hbnxb.2014.02.024.

http://www.hbnxb.net/CN/Y2014/V29/I2/127

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