KAMEL A. ABD-ELSALAM, FRANK SCHNIEDER, JIAN-RONG GUO
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引用次数: 18
摘要
我们改进了一种快速、廉价且不易污染的方法，将冷冻保存或新鲜菌丝体直接培养在96深孔板上。该方法便于通过非对称PCR (a -PCR)对单次提取的ssDNA真菌多样性进行评估。分离的镰刀菌DNA产率较高，在琼脂糖凝胶中加入10 μL的PCR产物，可观察到清晰的DNA条带。该程序可在不到4小时内完成，同时可处理96个样品。
A MODIFIED DNA EXTRACTION MINIPREPARATION PROTOCOL FOR FUSARIUM ISOLATES
Abstract We have modified a quick, inexpensive and less prone to contamination protocol by culturing the cryopreserved or fresh mycelium directly in 96-deepwell plate. The method facilitates concomitant assessment of ssDNA fungal diversity by asymmetric PCR (A-PCR) from a single extraction. The DNA yields from Fusarium spp. isolates was reasonably high, and a clear DNA band was frequently seen when 10 μL of the PCR product was run in agarose gel. The procedure can be completed in less than 4 h and 96 samples can be processed at the same time.
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