A rapid, Simple Genomic DNA Extraction from Yeasts for Large Scale Genetic Analyses

سال انتشار: 1397
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 503

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شناسه ملی سند علمی:

CIGS15_307

تاریخ نمایه سازی: 13 بهمن 1398

چکیده مقاله:

DNA extraction methods usually rely on time-consuming procedures that may involve handling hazardous compounds or using expensive kit. Here, we have developed a rapid genomic DNA extraction technique from yeasts and applied it for amplification of genomic DNA using gene specific primers. DNA extraction was evaluated at different stages from initial step to final pure DNA. In addition to final pure DNA of each sample which runs on an agarose gel to check the quantity and efficiency of whole yield, the extracts at different stages of extraction procedure were qualitatively examined and evaluated using gene amplification in a PCR reaction for minimal time consumption and lack of PCR inhibitors. The results showed that the earliest step for successful gene amplification was immediate after one hour incubation in 65 oC with extraction buffer. The obtained DNA was validated with different sets of primers for DNA amplification. The PCR products from each sliced bands were purified and sequenced. This evaluation showed that the presented method is a reliable, rapid and simple procedure with minimal equipment and resources requirements which is also environmentally friendly in absence of toxic chemicals for a cost efficient DNA extraction especially for large number of samples in less than one hour for sufficient genomic DNA for different purposes from yeasts for large scale genetic analyses including downstream PCR-based genetic analysis.

نویسندگان

Faezeh Ajorloo

Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), P.O. Box ۳۳۵۳۵۱۱۱, Tehran, Iran

Mohsen Vaez

Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), P.O. Box ۳۳۵۳۵۱۱۱, Tehran, Iran

Jafar Hemmat

Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), P.O. Box ۳۳۵۳۵۱۱۱, Tehran, Iran