摘要
目的寻找一种针对产孢少的丝状真菌简单、有效的DNA抽提方法。方法以红色毛癣菌和须癣毛癣菌为代表菌株,分别用改良后的氯化苄法及小量酵母DNA试剂盒(裂解酶+蛋白酶K)法抽提基因组DNA,并对所抽提DNA的质量及稳定性进行比较研究。结果氯化苄法抽提的皮肤癣菌DNA片段均>21 kb,每克菌丝体的DNA产量均>25μg。小量酵母基因组DNA试剂盒法抽提DNA,初步估测1．0×109／mL孢子菌丝混悬液的DNA得率仅约为0．6μg。采用氯化苄法抽提保存3年的皮肤癣菌基因组DNA仍能很好地适用于聚合酶链反应(PCR)扩增。结论采用氯化苄法抽提丝状真菌的DNA得率和纯度均较高,而且稳定性好,推荐用于产孢少、生长缓慢的丝状真菌DNA的抽提研究。
Objective To search for an effective DNA extraction method from filamentous fungi. Methods Improved extraction method using benzyl chloride was compared with Yeast DNA extraction Kit（Lyticase＋ Proteinase K） in extracting genomic DNA of Trichophyton rubrum and T. mentagrophytes in stability and quality. Results DNA extracted from per gram mycelia by improved benzyl chloride method was above 25μg with a molecular weight more than 21 kb. It also gave satisfactory results in molecular analysis even after being stored at -20 ℃ for 3 years. Whereas genomic fungal DNA yielded by Yeast DNA extraction Kit（Lyticase＋ Proteinase K） was only 0.6/μg per 1.0×10^9/mL. Conclusion Improved benzyl chloride method provides a rapid and effective procedure for extracting genomic DNA with high purity and high molecular weight from filamentous fungi.
出处
《上海医学》
CAS
CSCD
北大核心
2007年第1期14-17,共4页
Shanghai Medical Journal
关键词
丝状真菌
DNA
抽提
氯化苄
Filamentous fungi
DNA
Extract
Benzyl chloride
