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Optimization and comparative evaluation of nucleic acids extraction protocols

Author(s): Lucian Negura | George Popoiu | Mioara Matei | Doina Azoica | Anca Negura

Journal: Analele Ştiinţifice Ale Universităţii Alexandru Ioan Cuza din Iași,Sectiunea II A : Genetica si Biologie Moleculara
ISSN 1582-3571

Volume: 12;
Issue: 4;
Start page: 41;
Date: 2011;
Original page

Modern molecular applications have grown the need of biobanks, which contain DNA and RNA of high purety, quality, and quantity. Nucleic acid extraction methods have widely variate and evolve in time, from methods using toxic reagents to enzymatic protocols, and furthermore to DNA or RNA-binding polymers, separating membranes or highly eulogized commercially kits. In order to establish a constant, reproducible and ergonomic system in generating biobanks, we compared different available methods for the extraction of genomic DNA and total RNA, from peripheram blood or solid tumoral tissues. We evaluated the cost/effectiveness and time consumption of each method, tracking RNA/DNA quantity, quality and integrity. We imagined a “E-ratio” value to define these parameters, and a “NA- estimation” to integrate “E-ratio” with quality and integrity data.

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