一种适合于PCR扩增的真菌基因组DNA提取方法

RAPID EXTRACTION OF FUNGAL GENOMIC DNA FOR PCR AMPLIFICATION

以药用真菌灰树花和蛹虫草以及4种植物内生担子菌为材料,优化一种适用于PCR扩增的高质量基因组DNA提取方法。结果表明,采用改良SDS法提取药用真菌和植物内生担子菌基因组DNA的数量和质量都较为理想,A260/A280为1.8～1.9,DNA产量在110～170μg.g-1湿菌体。将提取的DNA作为模板PCR扩增rDNA ITS片段,扩增条带清晰,结果稳定、准确。该方法简便易行,成本低廉,适合富含蛋白质和多糖的真菌基因组DNA的提取。

An efficient method for extracting high quality genome DNA from two medicinal iungi and lour enctophytic fungi which had plenty of proteins, polysaccharides and many other chemical substances was optimized for PCR amplification. The results showed that quality and quantity of genomic DNA extraction from medicinal fungi and endophytic fungi with improved SDS method were perfect. The DNA purity was checked by analyzing the ra- tio of A260/A280 and 110 · 1701μg · g -1 DNA were obtained from every gram mycelia（wet weight）of different strains. They were used as the templates of PCR and the amplified bands were clear, stable and reliable. This technique was simple, convenient and inexpensive, which was especially adapted to extraction of fungal genomic DNA with abundant proteins and polysaccharides.