A Rapid Method of Preparing DNA Template of Filamentous Fungi for PCR Amplification

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.010

Abstract

Abstract:

This work aims to establish a novel method of extracting DNA template of filamentous fungi for PCR amplification with boiling-water bath and Fusarium oxysporum as raw material. The procedures of this method are as the following：exposing the fungal mycelia in certain volume of 50 mmol/L NaOH solution under boiling water bath, adding 1/10 volume of 1mol/L Tris-HCl(pH8.0)buffer, centrifuging it at 12 000 r/min for 10 min, and using the supernatant as the DNA template for PCR amplification. Results demonstrated that the efficiency of PCR amplification with the DNA template prepared by the method was high, the amplified bands were clear, and adaptability to various Taq DNA polymerases was strong；This indicated that the method was suitable for the preparation of DNA template of filamentous fungi. Conclusively, this method is economic, simple, rapid, safe and high-efficient, and can be utilized for high-throughput screening of filamentous fungal transformants and rapid identification of isolated strains.

Key words: filamentous fungi, Fusarium oxysporum, PCR amplification, Taq DNA polymerase

Luo Zhongqin, Cheng Lin, Zhang Xi, Chen Guohua. A Rapid Method of Preparing DNA Template of Filamentous Fungi for PCR Amplification[J]. Biotechnology Bulletin, 2015, 31(9): 79-83.

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