A rapid and efficient method of fungal genomic DNA extraction, suitable for PCR based molecular methods

By Sutar Aamir S, Baghela. 2015

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We report a rapid and efficient method of genomic DNA extraction from filamentous fungi with high-throughput potential. The method involves disruption of fungal cell by bead beating in a homogenizer, followed by RNase treatment, phenol: chloroform: Isoamyl alcohol extraction and precipitation with isopropanol. The method does not involve any enzymatic digestion and it can be completed within 2.5 hours. The method yielded good quality and quantity (60 μg – 230 μg/200 mg of wet fungal mass) of the DNA. Being a closed system of gDNA extraction, our method has been found to be useful in avoiding the laboratory borne contamination during DNA extraction. The extracted DNA was found to be suitable for PCR based molecular methods like single and multicopy gene amplification and RAPD analysis.

Subfields: Cell Biology, Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/0705bcca4fbcfbd41ebc72eb9747dba463b024fa