球形孢子丝菌DNA提取方法的比较

中国真菌学杂志　 2015, Vol. 10 　Issue (4): 216-219.

球形孢子丝菌DNA提取方法的比较

石滢, 崔岩, 赵莉佩, 宋洋, 李珊山

吉林大学第一医院皮肤性病科, 长春 130021

收稿日期:2015-05-25 出版日期:2015-08-28 发布日期:2015-08-28
通讯作者: 李珊山,E-mail:shansalee@163.com E-mail:shansalee@163.com
作者简介:石滢,女(汉族),硕士研究生在读.E-mail:442387194@qq.com
基金资助:
国家自然科学基金(81171509);吉林省科技厅科技创新人才培育计划(20150520039JH)

Comparison of DNA extraction methods from Sporothrix globosa

SHI Ying, CUI Yan, ZHAO Li-pei, SONG Yang, LI Shan-shan

Department of Dermatology and Venereology, First Hospital of Jilin University, Changchun 130021, China

Received:2015-05-25 Online:2015-08-28 Published:2015-08-28

摘要/Abstract

摘要：

目的获得一种高效、快速、简便的提取球形孢子丝菌DNA的方法。方法以不同地区来源的球形孢子丝菌为实验材料,采用CTAB法、异硫氰酸胍法、试剂盒法、碱裂解法4种方法分别提取球形孢子丝菌DNA。通过紫外吸光度法测定DNA的浓度和纯度,琼脂糖凝胶电泳、PCR扩增评价DNA的质量。结果虽然不同方法均可得到球形孢子丝菌的DNA,但碱裂解法获得DNA的浓度和纯度明显优于其他3种方法。利用钙调蛋白基因的引物对DNA进行PCR扩增后,各样本均出现阳性条带,且条带亮度较好。结论碱裂解法是获得高质量球形孢子丝菌DNA的理想方法。

关键词: 球形孢子丝菌, 碱裂解法, CTAB法, 异硫氰酸胍法, DNA提取

Abstract:

Objective To find a rapid, effective and easy-to-perform DNA extraction protocol from Sporothrix globosa.Methods Four methods, such as CTAB-extraction, guanidine thiocyanate boiling-extraction, alkaline lysis-extraction, DNA extraction kit, were compared based on Sporothrix globosa isolates from different regions.The purity and yield of the DNA were determined by ultraviolet absorption spectrum.The DNA quality was evaluated with agarose gel electrophoresis and PCR amplification.Results It showed that all methods could get DNA, but alkaline lysis-extraction was better than others for the purity and yield of the DNA.Based on degenerated primers of fragments of the calmodulin genes for PCR, we found all samples showed bright stripe.Conclusion Alkaline lysis-extraction is the most suitable method for extracting high quality DNA from Sporothrix globosa.

Key words: Sporothrix globosa, Alkaline lysis-extraction, CTAB-extraction, Guanidine thiocyanate boiling-extraction, DNA extration

中图分类号:

R379.9

石滢, 崔岩, 赵莉佩, 宋洋, 李珊山. 球形孢子丝菌DNA提取方法的比较[J]. 中国真菌学杂志, 2015, 10(4): 216-219.

SHI Ying, CUI Yan, ZHAO Li-pei, SONG Yang, LI Shan-shan. Comparison of DNA extraction methods from Sporothrix globosa[J]. Chinese Journal of Mycology, 2015, 10(4): 216-219.

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