一种快速提取丝状真菌染色体DNA的方法

A rapid method for preparation of filamentous fungi chromosome DNA

介绍了一种适用于丝状真菌染色体DNA大片段的快速提取方法,该方法以(100mM Tris,100mM NaG l,50mMEDTA-Na22%SDS,pH值9.0)为提取液,经石英砂研磨破壁。应用该方法成功地提取了粗糙脉胞菌(Neurospora crassa)、米曲霉(Aspergillus oryzae)、产黄青霉(Penicillium chrysogenum)和头孢霉菌(Cep-halosporiumsp.)等4种不同丝状真菌的染色体DNA大片段,且所提DNA片段均大于20kb,可直接用于限制性酶切、PCR等分子生物学研究。

A rapid method for preparation of fungal chromosome DNA was introduced.In this method,the solution containing 100mM Tris,100mM NaCl,50mM EDTA-Na2,and 2% SDS,pH 9.0 was used as the extraction buffer by crashing cell walls of fungi with quartz sand.As the results,the fragments with high qualitive chromosomal DNA were obtained from 5 species of fungi Neurospora crassa,Aspergillus oryzae,Penicillium chrysogenum,Cephalosporium sp..And the lengths of all chromosomal DNA fragments from fungi were longer than 20 kb in nucleotides and were able to be used directly for either digestion with restriction endo-enzyme or PCR analysis.