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12 \ 09 \ 2018

RNA Isolation Kits (Technical note)

    Eva Bace  Barbora Dvorakova,                

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Technical information describing the performance characteristics and isolation efficacy for the BioVendor RNA Isolation kits is provided in this note. These characteristics are compared with commonly used commercial kits.

 

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Performance Characteristics of BioVendor RNA Isolation Kits


Introduction

Sample collection and processing is crucial for correct miRNA quantification. To address this problem, BioVendor has launched two RNA isolation kits, the RNA Isolation kit (RIK001) for whole blood, cell cultures and tissues and RNA Isolation kit Plasma/Serum (RIK002) for plasma and serum samples.
For the description and protocols see www.biovendor.com/mirna.  

Experimental design

All samples were collected in PAXgene Blood RNA tubes (PreAnalytiX). BioVendor RNA Isolation kit Plasma/Serum (RIK002) was used for isolation of total RNA from EDTA plasma and serum; the RNA Isolation kit (RIK001) was used for peripheral blood mononuclear cells (PBMC) and whole blood. RNA was isolated from the same samples using competitor kits and extracted with their protocol using chloroform/phenol extraction. During all isolations, rDNase was added. Total RNA concentrations were measured in a NanoQuant infinite M200Pro reader (Tecan), hsa-miR-16-5p was determined specifically by a miRNA enzyme immunoassay kit (hsa-miR-16-5p miREA RDM0008H, BioVendor) and a qPCR kit (miRCURY LNA™ Universal RT microRNA PCR, Exiqon). 

Results

BioVendor RNA Isolation kits were used for isolation of total RNA from all matrices included in the experiment. Isolated high-quality RNA fractions contained target microRNA hsa-miR-16-5p suitable for the miREIA and qPCR determination. hsa-miR-16-5p levels determined by miREIA and qPCR were comparable in all RNA fractions isolated by BioVendor and competitor isolation kits

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Conclusions

Our experiment proved that BioVendor RNA isolation kits offer a solution for effective and efficient extraction of high-quality RNA, including miRNA. The kits fulfilled rigorous requirements for downstream applications, the quality of RNA fractions isolated with BioVendor kits were fully comparable to those obtained with the competitor kits. The main advantage of BioVendor kits over the competitor was no need of chloroform/phenol extraction in the isolation procedure. rDNase is added during isolation to obtain high purity RNA fractions and avoid possible DNA interference in qPCR reactions. BioVendor kits contain the rDNase reagent while the competitor kits do not. Sample purification by BioVendor RNA isolation kits proved to be optimal for subsequent miRNA quantification by miREIA immunoassays.

 

 


 


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