Lyophilized human cells stored at room temperature preserve multiple RNA species at excellent quality for RNA sequencing

Oncotarget
Lilla OzgyinBalint Laszlo Balint

Abstract

Biobanks operating at ambient temperatures would dramatically reduce the costs associated with standard cryogenic storage. In the present study, we used lyophilization to stabilize unfractionated human cells in a dried state at room temperature and tested the yield and integrity of the isolated RNA by microfluidic electrophoresis, RT-qPCR and RNA sequencing. RNA yields and integrity measures were not reduced for lyophilized cells (unstored, stored for two weeks or stored for two months) compared to their paired controls. The abundance of the selected mRNAs with various expression levels, as well as enhancer-associated RNAs and cancer biomarker long non-coding RNAs (MALAT1, GAS5 and TUG1), were not significantly different between the two groups as assessed by RT-qPCR. RNA sequencing data of three lyophilized samples stored for two weeks at room temperature revealed a high degree of similarity with their paired controls in terms of the RNA biotype distribution, cumulative gene diversity, gene body read coverage and per base mismatch rate. Among the 28 differentially expressed genes transcriptional regulators, as well as certain transcript properties suggestive of a residual active decay mechanism were enriched. Our study suggests...Continue Reading

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Citations

Jun 12, 2020·PloS One·Raffaele BrognaWillem F Wolkers

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Datasets Mentioned

BETA
GM12872
GM12873
GM12878
GSM1872887
GSM1480323
GSE106344

Methods Mentioned

BETA
biopsies
RNA-Seq
PCR
electrophoresis
electrophoretic methods
using
flow
immunoprecipitation
Assay
ChIP-seq

Software Mentioned

ARED
bcl2fastq Conversion
Cuffdiff
GraphPad
Picard
TopHat
AccuGENE
GraphPad Prism
SAMtools
MarkDuplicates

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