A simple method to extract filamentous fungal genomic DNA

Subject Areas : دو فصلنامه تحقیقات بیماریهای گیاهی

Saeed Ghafoori Harat ¹ (دانش آموخته کارشناسی ارشد، گروه بیماری شناسی گیاهی، واحد مرودشت، دانشگاه آزاد اسلامی، مرودشت، ایران.)
Tahereh Basirnia ² (استادیار، گروه بیماری شناسی گیاهی، واحد مرودشت، دانشگاه آزاد اسلامی، مرودشت، ایران.)
Mohammad Reza Moosavi ³ (استادیار، گروه بیماری شناسی گیاهی، واحد مرودشت، دانشگاه آزاد اسلامی، مرودشت، ایران.)

Received: 2016-02-01 Accepted : 2016-02-01 Published : 2016-05-21

Keywords: C-TAB, ITS, Antagonist, DNA extraction, filamentous fungi, grinder,

Abstract :

Today, the application of molecular methods is a common and necessary part of many routine researches. DNA extraction is the foundation for most of such research studies. As a result, it is of great significance to discover and select proper techniques that are productive, inexpensive, straightforward, safe and rapid at the same time. Agricultural research and experiments are no exception in this respect. C-TAB, phenol-chloroform and DNA extraction kits are conventional techniques in agriculture for extracting DNA from filamentous fungi. Each of these methods has its advantages and disadvantages. In this study, we have developed a novel, straightforward and effective method to extract the DNA from antagonistic-, saprophytic- and plant pathogenic-filamentous fungi. This method is a composition of the aforementioned techniques and is used to avoid the application of liquid nitrogen and phenol. It is called squish with grinder and is a safer, more affordable and quicker method that results in relatively pure DNAs. The purity of obtained DNA was assessed by spectrophotometry. The ITS region and β-tubuline gene was amplified by PCR. The results of this assessment justify this method as a practical and reliable technique to extract filamentous fungal genomic DNA.

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