Simultaneous isolation of mRNA and native protein from minute samples of cells

Research output: Contribution to journalJournal articleResearchpeer-review

Precious biological samples often lack a sufficient number of cells for multiple procedures, such as extraction of mRNA while maintaining protein in a non-denatured state suitable for subsequent characterization. Here we present a new method for the simultaneous purification of mRNA and native proteins from samples containing small numbers of cells. Our approach utilizes oligodeoxythymidylate [oligo(dT)25]-coated paramagnetic beads in an optimized reaction buffer to isolate mRNA comparable in quantity and quality to mRNA isolated with existing methods, while maintaining the proteins in their native state for traditional protein assays. We validated the procedure using neonatal rat ovaries and small numbers of human granulosa cells, demonstrating the extraction of mRNA suitable for gene expression analysis with simultaneous isolation of native proteins suitable for downstream characterization using different protein assays.

Original languageEnglish
JournalBioTechniques
Volume56
Issue number5
Pages (from-to)229-237
Number of pages9
ISSN0736-6205
DOIs
Publication statusPublished - 2014

    Research areas

  • Animals, Caspase 3, Female, Granulosa Cells, Humans, Inhibins, L-Lactate Dehydrogenase, Molecular Biology, Ovary, Protein Denaturation, Proteins, RNA, Messenger, Rats, Wistar, Reproducibility of Results, Ribosomal Proteins, Transcriptome

ID: 138421228