Optimized microRNA purification from TRIzol-treated plasma

BMC Genomics
Janice DuyTimothy D Minogue

Abstract

MicroRNAs (miRNAs) represent new and potentially informative diagnostic targets for disease detection and prognosis. However, little work exists documenting the effect of TRIzol, a common viral inactivation and nucleic acid extraction reagent, on miRNA purification. Here, we developed an optimized protocol for miRNA extraction from plasma samples by evaluating five different RNA extraction kits, TRIzol phase separation, purification additives, and initial plasma sample volume. This method was then used for downstream profiling of plasma miRNAs found in archived samples from one nonhuman primate (NHP) experimentally challenged with Ebola virus by the aerosol route. Comparison of real-time RT-PCR results for spiked-in and endogenous miRNA sequences determined extraction efficiencies from five different RNA purification kits. These experiments showed that 50 μL plasma processed using the QIAGEN miRNeasy Mini Kit with 5 μg of glycogen as a co-precipitant yielded the highest recovery of endogenous miRNAs. Using this optimized protocol, miRNAs from archived plasma samples of one rhesus macaque challenged with aerosolized Ebola virus was profiled using a targeted real-time PCR array. A total of 519 of the 752 unique miRNAs assayed wer...Continue Reading

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Citations

Jun 10, 2015·Journal of Zhejiang University. Science. B·Xiao-lu JinJian-xin Liu
Apr 22, 2016·Scientific Reports·Janice DuyTimothy D Minogue
Jul 7, 2016·The International Journal of Neuroscience·Meire Silva BatistelaRicardo Lehtonen Rodrigues de Souza
Dec 9, 2017·Chronobiology International·Diego de Siqueira FigueredoTiago Gomes de Andrade
Apr 25, 2018·Scientific Reports·Janice DuyTimothy D Minogue
May 12, 2019·Veterinary Clinical Pathology·Candice P Chu, Mary B Nabity
Oct 17, 2017·Analytical and Bioanalytical Chemistry·Luis A JimenezWenwan Zhong
Jul 10, 2020·Journal of Assisted Reproduction and Genetics·Maryam Qasemi, Fardin Amidi
Jan 23, 2020·Scientific Reports·Kathryn WrightKarren M Plain
Mar 14, 2019·Scientific Reports·Peter AndrovicLukas Valihrach
Oct 23, 2019·Molecular Aspects of Medicine·Lukas ValihrachMikael Kubista

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Methods Mentioned

BETA
PCR
blood collection

Software Mentioned

miRNome
Roche LightCycler 480
R
Exiqon
GenEx

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