کارآیی روش‌های استخراج RNA از برگ گیاه دارویی سرخارگل
محورهای موضوعی : بوم شناسی گیاهان زراعی
1 - بخش تحقیقات سلولی مولکولی، مدیریت بیوتکنولوژی کشاورزی منطقه مرکزی کشور- اصفهان، پژوهشکده بیوتکنولوژی کشاورزی ایران، سازمان تحقیقات،
2 - بخش تحقیقات ژنومیکس، مدیریت بیوتکنولوژی کشاورزی منطقه مرکزی کشور- اصفهان، پژوهشکده بیوتکنولوژی کشاورزی ایران، سازمان تحقیقات، آموزش و ترویج کشاورزی، تهران
کلید واژه: روش لیتیم کلراید, روش لیتیم کلراید/فنل, کیت RNAX-Plus, روش فنل/SDS,
چکیده مقاله :
سرخارگل گیاه دارویی ارزشمند حاوی سطوح بالایی از متابولیت های ثانویه می باشد که تا کنون روش مناسبی جهت استخراج RNA کل از آن معرفی نشده است. در پژوهش حاضر چهار روش استخراج RNA شامل روش لیتیم کلراید، کیت RNAX-Plus، فنل/SDS و لیتیم کلراید/فنل مورد بررسی قرار گرفت. به منظور تأیید کیفیت و خلوص RNA از روش اسپکتروفتومتری و الکتروفورز روی ژل آگارز 5/1% استفاده شد. نسبت بالای جذب محلول RNA در طول‌موج 260 نانومتر به طول موج 280 و 230 نانومتر، مشاهده RNA های ریبوزمی و عدم مشاهده اسمیر روی ژل به عنوان معیاری برای تعیین کیفیت نمونه‌ها در نظر گرفته شد. بالاترین کیفیت RNA حاوی 9/2 میکروگرم بر میکرولیتر در 100 میلی‌گرم برگ تازه در روش فنل/SDS و کمترین کیفیت RNA حاوی 2/1 میکروگرم در 100 میلی‌گرم برگ تازه در روش لیتیم کلراید/فنل مشاهده شد. بنابراین، روش فنل/SDS به عنوان مناسب ترین روش برای استخراج RNA برگ گیاه سرخارگل معرفی می گردد
Coneflower is a valuable medicinal plant containing high levels of secondary metabolites, but no suitable method for extraction of the entire RNA has been introduced yet. In present study, four methods of RNA extraction including lithium chloride, RNAX-Plus kit, phenol/SDS and lithium chloride/phenol were investigated. For confirming the quality and purity of RNA, spectrophotometer and agarose gel electrophoresis of 1.5% were used. The high absorption ratio of RNA solution at a wavelength of 260 nM to 280 and 230 nM, observation of ribosomal RNAs and the lack of smear on the gel were considered as criteria for determining samples quality. The highest and the lowest qualities of RNA contained 2.9 μg/100 mg of fresh leaves by phenol/SDS method and 1/2 μg/100 mg of fresh leaves by lithium chloride/phenol method, respectively. Therefore, phenol/SDS method is introducing as the most suitable method for RNA extraction from coneflower.
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