Evaluation of the efficacy of various reagents in improving microRNA extraction

Annals of Clinical Biochemistry
Arizumi KikuchiKenichi Nonaka

Abstract

MicroRNA has received considerable attention in the clinical context, and attempts are being made to use microRNA in clinical diagnosis. However, adequate quantities of microRNA required for analysis are challenging to isolate. We tested the effect of various reagents in improving microRNA extraction and compared their efficacy to that of a commercially available extraction kit (HighPure miRNA isolation kit, Roche). We used the synthetic oligonucleotide miR-21 and formalin-fixed, paraffin-embedded (FFPE) tissue sections from colon cancer samples ( n = 10). We tested increasing volumes (100-600  μL) of 1,4-dioxane, 2-butanol, 2-propanol, acetonitrile, polyethylene glycol (PEG) 600, PEG 1000, PEG 1540, PEG 2000, tetraethylene glycol dimethyl ether (TDE), and tetrahydrofuran, instead of the binding enhancer solution provided in the kit. MiR-21 analysis was performed via stem-loop RT-qPCR using Universal ProbeLibrary probe (Roche). The optimum amount of each enhancement solution was 200-500  μL. We obtained ΔCp values of optimum additional volume for each solution from 1.04 to 2.50 and compared these with those obtained using the commercially available kit. PEG 1540 and 2000 produced superior reactivity with minimal addition. For F...Continue Reading

References

Mar 1, 1990·Journal of Clinical Microbiology·R BoomJ van der Noordaa
Sep 25, 2001·The Journal of Pathology·F LewisP Quirke
Oct 28, 2005·The New England Journal of Medicine·George Adrian CalinCarlo M Croce
Oct 3, 2006·Developmental Cell·Wigard P Kloosterman, Ronald H A Plasterk
Dec 23, 2009·The International Journal of Biochemistry & Cell Biology·William C S Cho
Sep 18, 2010·Clinical Chemistry·Jessica A WeberKai Wang
Oct 5, 2010·Nature Reviews. Drug Discovery·Ramiro GarzonCarlo M Croce
Oct 8, 2013·Nature Structural & Molecular Biology·Anders JacobsenChris Sander
Nov 22, 2013·Wiley Interdisciplinary Reviews. RNA·Hongwei LiangXi Chen
Feb 5, 2014·Nature Reviews. Clinical Oncology·Heidi SchwarzenbachKlaus Pantel
Feb 19, 2014·Journal of Cellular and Molecular Medicine·Leni MoldovanMelissa Piper
Jun 18, 2014·EMBO Molecular Medicine·Eva van Rooij, Sakari Kauppinen
Nov 14, 2014·Clinical Chemistry·Kenneth W Witwer
Dec 17, 2014·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Ioana Berindan-Neagoe, George A Calin
Feb 20, 2015·TheScientificWorldJournal·Jin LinXiao-Zhong Wang
Oct 30, 2015·Journal of Clinical Medicine·Shigeshi OnoDave S B Hoon

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biopsy
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