Investigation of MicroRNA expression in experimental glaucoma

Hari Jayaram, Diana C. Lozano, Elaine Johnson, John Morrison

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

MicroRNAs are small, endogenous noncoding RNAs that modulate post-transcriptional gene expression. Recent evidence suggests that they may have a potential role in the regulation of the complex biological responses that develop in response to elevated intraocular pressure. However, contemporary microRNA assay techniques (e.g., microarrays and next-generation sequencing) typically require large amounts of RNA template that are often times difficult to obtain from glaucomatous tissue. We describe in detail an experimental protocol utilizing targeted pre-amplification and low-density polymerase chain reaction arrays to circumvent this hurdle. This approach optimizes the simultaneous high-throughput screening of small tissue samples, such as the rodent optic nerve head, for up to 754 microRNA probes while also providing an opportunity for subsequent confirmatory reactions of technical or biological replicates.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages287-297
Number of pages11
Volume1695
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1695
ISSN (Print)1064-3745

Keywords

  • Gene expression
  • MicroRNA
  • MicroRNA array card
  • Optic nerve head
  • PCR
  • Post-transcriptional silencing
  • Quantitative PCR
  • Taqman

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Jayaram, H., Lozano, D. C., Johnson, E., & Morrison, J. (2018). Investigation of MicroRNA expression in experimental glaucoma. In Methods in Molecular Biology (Vol. 1695, pp. 287-297). (Methods in Molecular Biology; Vol. 1695). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7407-8_19