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

عنوان مقاله: A rapid, Simple Genomic DNA Extraction from Yeasts for Large Scale Genetic Analyses
شناسه ملی مقاله: CIGS15_307
منتشر شده در سومین کنگره بین المللی و پانزدهمین کنگره ملی ژنتیک ایران در سال 1397
مشخصات نویسندگان مقاله:

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

خلاصه 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.

صفحه اختصاصی مقاله و دریافت فایل کامل: https://civilica.com/doc/983862/