Biology Reference
In-Depth Information
Chapter 10
MicroRNA Expression Profi ling of Rat Blood and Brain
Tissues: TaqMan Real-Time PCR MicroRNA Assays
Da-Zhi Liu , Bradley P. Ander , and Frank R. Sharp
Abstract
Ischemic and hemorrhagic stroke alters microRNA (miRNA) expression in animals and patients. The
recent discovery of miRNAs as key regulators of gene function has introduced a new level and mechanism
of gene regulation, with estimates that greater than one-third of all human genes may be regulated by
miRNAs. TaqMan real-time PCR is a specifi c, sensitive, and accurate method for assessing miRNA expres-
sion, allowing direct analysis in a single cell without the need for nucleic acid purifi cation. This chapter
introduces a brief history of Taqman real-time PCR, required materials and procedures, software, and
statistical analysis.
Key words:
MicroRNA, Stroke, Stem-loop reverse transcription, TaqMan, Real-time PCR
1. Introduction
Ischemic and hemorrhagic stroke alters microRNA (miRNA)
expression in rat blood and brain (
1-3
) and in the blood of humans
(
4
). The recent discovery of miRNAs, which are endogenous, non-
coding, single-stranded RNA molecules of 19-25 nucleotides in
length, has introduced a new level and mechanism of gene regulation.
Though there exist only hundreds of miRNAs, each of them can
potentially regulate hundreds of target genes. Current predictions
estimate that miRNA may regulate greater than one-third of all
human genes. Therefore, miRNA expression studies are attracting
more and more attention for diagnosis and prognosis of illness, as
well as giving insight into the understanding of the pathogenesis of
many medical and neurological diseases.
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