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بررسی مقایسه­ ای سه روش استخراج DNA ژنومی از نمونه­ های تازه و گیاکده ­ای برگ بومادران کوتاه دشتی

نویسندگان

عذرا صبورا 1
مریم امیری راد 1
عزت عسگرانی 1
طیبه رجبیان 2

1 دانشگاه الزهرا

2 دانشگاه شاهد

https://doi.org/10.29252/nbr.5.4.458
چکیده
استخراج DNA از بافت­های گیاهی اغلب مشکلاتی را فراهم می­سازد. بطورمثال عدم حذف متابولیت­های ثانویه نظیر ترکیبات فنلی گیاهان معطر و دارویی، باعث بروز خطا در نتایج کاربرد DNA استخراج شده طی آزمایش های مولکولی می­شود. بومادران (Achillea wilhelmsii)، گیاهی دارویی متعلق به تیره کاسنیان (Asteraceae) و انحصاری ایران است که اطلاعات کمی در رابطه با ژنوم آن وجود دارد. بدین جهت بهینه­سازی روش­های استخراج برای تهیه DNA با کمیت و کیفیت مناسب ضروری به نظر می­رسد. در این مطالعه دو روش سنتی استخراج DNA ژنومی از برگ گیاه بومادران (با استفاده از بافت تازه و گیاکده­ای) و یک روش دیگر با استفاده از کیت استخراج تجاری MAGNANTM مورد مقایسه قرار گرفته است. نتایج نشان داد که استخراج DNA از برگ­های تازه طبق دستورالعمل Khanuja و همکاران (1999) به دلیل افزایش مقدار DNA استخراج شده و کاهش آلودگی ترکیبات مزاحم مثل RNA، پلی­ساکاریدها، پروتئین­ها و متابولیت­های ثانوی بهتر از دو روش دیگر بود. ارزیابی کیفیت DNA استخراج شده از نمونه­های گیاکده­ای نشان داد که علیرغم مقدار بالای DNA (50-10 برابر نسبت به دو روش دیگر)، کیفیت عصاره روی ژل آگارز بخاطر وجود کشیدگی و شکستگی مولکول DNA پایین بود. این امر می­تواند استفاده از عصاره فوق را در آزمایشهای مولکولی با محدودیت روبرو سازد. کاربرد کیت استخراج MAGNANTM گرچه از نظر کاهش زمان و هزینه استخراج و کاهش آلودگی به پروتئین و RNA مناسب بود، اما نیازمند اصلاح فرایند تخلیص برای گیاه بومادران جهت افزایش کمیت DNA حاصل است.


کلیدواژه‌ها

استخراج ژنومی
بومادران
پلی فنل ها
نانوذرات مغناطیسی
نمونه خشک

عنوان مقاله English

Comparison study of three methods for genomic DNA extraction from fresh and herbarium leaf specimens of Achillea wilhelmsii C. Koch

نویسندگان English

Azra Saboora 1
Maryam Amiri Rad 1
Ezat Asgarani 1
Tayebeh Radjabian 2
1 alzahra university
2 shahed university
چکیده English

DNA extraction from plant tissues often causes most problems. For example, unsuccessful removal secondary metabolites during extraction, such as phenolic compounds in aromatic and medicinal plants, cause to some mistakes in result of molecular experiments by using of the extracted DNA. Achillea wilhelmsii is a medicinal plant belong to Asteraceae family and native to Iran, there is little information about genomic data in this plant. Therefore, optimizing of the DNA extraction methods for obtaining suitable quality and quantity yield is necessary. In this study, two traditional DNA extraction methods (using fresh and herbarium leaf samples) and commercial DNA kit MAGNANTM in Yarrow have been compared. Results showed that DNA extracted from fresh leaves of yarrow according to Khanuja et al. (1999) was better than the other methods which mentioned in this research, because of the increased amount of extracted DNA and reduced harmful compounds such as RNA, polysaccharides, protein and secondary metabolites. To evaluate the quality of extracted DNA from herbarium specimens showed that, in spite of the high yielding DNA (10-50 fold ratio to the other methods), quality of this extract was low on agarose gel because of smear and broken down the molecules. It can be considered as a limitation of the extract in molecular experiments. Procedure of MAGNANTM DNA kit was appropriate for reducing time and cost of the extraction as well as low contamination to protein and RNA, but the process needs some modifications for yarrow to increase the amount of extracted DNA.

کلیدواژه‌ها English

Achillea wilhelmsii
genomic DNA extraction
herbarium samples
Magnetic Nanoparticle
polyphenols
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  • تاریخ دریافت 18 خرداد 1405
  • تاریخ انتشار 18 خرداد 1405

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