Investigation of MicroRNA Expression in Experimental Glaucoma
| Autor: | Hari Jayaram, Elaine C. Johnson, Diana C Lozano, John C. Morrison |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Regulation of gene expression RNA Computational biology Biology law.invention Gene expression profiling 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Real-time polymerase chain reaction law Gene expression microRNA 030221 ophthalmology & optometry DNA microarray Polymerase chain reaction |
| Zdroj: | Glaucoma ISBN: 9781493974061 |
| DOI: | 10.1007/978-1-4939-7407-8_19 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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