Elimination of heparin interference during microarray processing of fresh and biobank-archived blood samples

Environmental and Molecular Mutagenesis
Dennie G A J HebelsJos C S Kleinjans

Abstract

In the context of environmental health research, biobank blood samples have recently been identified as suitable for high-throughput omics analyses enabling the identification of new biomarkers of exposure and disease. However, blood samples containing the anti-coagulant heparin could complicate transcriptomic analysis because heparin may inhibit RNA polymerase causing inefficient cRNA synthesis and fluorophore labelling. We investigated the inhibitory effect of heparin and the influence of storage conditions (0 or 3 hr bench times, storage at room temperature or -80°C) on fluorophore labelling in heparinized fresh human buffy coat and whole blood biobank samples during the mRNA work-up protocol for microarray analysis. Subsequently, we removed heparin by lithium chloride (LiCl) treatment and performed a quality control analysis of LiCl-treated biobank sample microarrays to prove their suitability for downstream data analysis. Both fresh and biobank samples experienced varying degrees of heparin-induced inhibition of fluorophore labelling, making most samples unusable for microarray analysis. RNA derived from EDTA and citrate blood was not inhibited. No effect of bench time was observed but room temperature storage gave slightl...Continue Reading

References

Sep 1, 1992·Analytical Biochemistry·J E CoadC E Litz
Apr 1, 1991·Journal of Clinical Microbiology·M HolodniyT C Merigan
Aug 25, 1990·Nucleic Acids Research·M Goulian, C J Heard
Jan 1, 1987·Methods in Enzymology·D M Wallace
Dec 1, 1967·European Journal of Biochemistry·G WalterE Fuchs
Jun 11, 1994·Lancet·J SatsangiJ I Bell
May 14, 1999·Journal of Clinical Laboratory Analysis·M YokotaI Tsuda
Jun 3, 2000·Transplant International : Official Journal of the European Society for Organ Transplantation·X BaiM Liu
Nov 8, 2002·Mathematical Biosciences·C Connell McCluskey
Dec 20, 2003·BioTechniques·Mary Lynn JohnsonDale A Redmer
Sep 30, 2008·Toxicology Letters·Heidrun Ellinger-ZiegelbauerHans-Juergen Ahr
Aug 29, 2009·American Journal of Epidemiology·Leda ChatziManolis Kogevinas
Apr 13, 2010·Mutation Research·Cliona M McHaleMartyn T Smith
Dec 21, 2013·The Journal of Physical Chemistry. B·Consuelo N BeecherCynthia K Larive

Citations

Jan 6, 2017·Vector Borne and Zoonotic Diseases·Cornelia SilaghiJ Stephen Dumler
May 17, 2015·Clinical Chemistry·Christina Ellervik, Jim Vaught
May 25, 2016·Annual Review of Nutrition·Joëlle V FritzPaul Wilmes
Apr 4, 2021·Animals : an Open Access Journal From MDPI·Seungmee LeeSarah E Taylor

Related Concepts

Blood Banks
Blood Chemical Analysis
Fluorochromes
Liquaemin
RNA
Lithium Chloride
Microarray Analysis
Biological Markers
Blood
Environment

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