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Extensive studies have been conducted on the identification,
biogenesis, and processing of microRNA (miRNA) as well as research
on the exact mechanism by which miRNAs bring about translational
silencing of their targets. In addition, numerous publications
point to an important role of miRNAs in development, reprogramming,
epigenetics, pathogenesis of cancer, oncogenes and tumor
suppressors, biomarkers of various disease onset, and regulation of
adipogenesis and obesity; yet many questions still remain. In
MicroRNA Protocols, Second Edition, experts in the field provide
up-to-date coverage of this diverse area of study. The specific
chapters of this edition are related to the analysis of miRNA,
targets and expression profiling, various methods to determine its
regulation of gene expression, the preparation and isolation of
miRNAs in specific tissues, its detection in the saliva, and
potential application in cosmetics, wound healing, and prostate
cancer. Written in the highly successful Methods in Molecular
Biology (TM) series format, chapters include introductions to their
respective topics, lists of the necessary materials and reagents,
step-by-step, readily reproducible laboratory protocols, and tips
on troubleshooting and known pitfalls. Fully updated and easy to
use, MicroRNA Protocols, Second Edition aims to stimulate readers
to explore diverse ways to understand the mechanisms in which
miRNAs facilitate the molecular aspects of not only biomedical
research but also a wide range of other research fields.
Extensive studies have been conducted on the identification,
biogenesis, and processing of microRNA (miRNA) as well as research
on the exact mechanism by which miRNAs bring about translational
silencing of their targets. In addition, numerous publications
point to an important role of miRNAs in development, reprogramming,
epigenetics, pathogenesis of cancer, oncogenes and tumor
suppressors, biomarkers of various disease onset, and regulation of
adipogenesis and obesity; yet many questions still remain. In
MicroRNA Protocols, Second Edition, experts in the field provide
up-to-date coverage of this diverse area of study. The specific
chapters of this edition are related to the analysis of miRNA,
targets and expression profiling, various methods to determine its
regulation of gene expression, the preparation and isolation of
miRNAs in specific tissues, its detection in the saliva, and
potential application in cosmetics, wound healing, and prostate
cancer. Written in the highly successful Methods in Molecular
Biology trademark] series format, chapters include introductions to
their respective topics, lists of the necessary materials and
reagents, step-by-step, readily reproducible laboratory protocols,
and tips on troubleshooting and known pitfalls. Fully updated and
easy to use, MicroRNA Protocols, Second Edition aims to stimulate
readers to explore diverse ways to understand the mechanisms in
which miRNAs facilitate the molecular aspects of not only
biomedical research but also a wide range of other research fields.
Since its invention and subsequent development nearly 20 years ago,
po- merase chain reaction (PCR) has been extensively utilized to
identify numerous gene probes in vitro and in vivo. However,
attempts to generate complete and full-length complementary cDNA
libraries were, for the most part, fruitless and remained elusive
until the last decade, when simple and rapid methods were
developed. With current decoding and potential application of human
genome information to genechips, there are urgent needs for
identification of functional significance of these decoded gene
sequences. Inherent in bringing these app- cations to fruition is
the need to generate a complete and full-length cDNA library for
potential functional assays of specific gene sequences. Generation
of cDNA Libraries: Methods and Protocols serves as a laboratory
manual on the evolution of generation of cDNA libraries, covering
both ba- ground information and step-by-step practical laboratory
recipes for which p- tocols, reagents, operational tips,
instrumentation, and other requirements are detailed. The first
chapter of the book is an overview of the basics of generating cDNA
libraries, which include the following: (a) the definition of a
cDNA library, (b) different kinds of cDNA libraries, (c)
differences between methods for cDNA library generation using
conventional approaches and novel stra- gies, including reverse
generation of RNA repertoires from cDNA libraries, and (d) the
quality of cDNA libraries.
MicroRNA Protocols provides diverse, novel, and useful descriptions
of miRNAs in several species, including plants, worms, flies, fish,
chicks, mice, and humans. These include some useful adaptations and
applications that could be relevant to the wider research community
who are already familiar with the identification of miRNAs. This
volume will stimulate the reader to explore diverse ways to
understanding the mechanism in which miRNAs facilitate the
molecular aspects of the biomedical research.
Nearly 97% of the human genome is the non-coding DNA, which
varies from one species to another, and changes in these sequences
are frequently noticed to manifest clinical and circumstantial
malfunction. Numerous non-protein-coding genes are recently found
to encode microRNAs, which are responsible for RNA-mediated gene
silencing through RNA interference (RNAi)-like pathways. MicroRNAs
(miRNAs), small single-stranded 17-25 nucleotide RNAs capable of
interfering with intracellular messenger RNAs (mRNAs) that contain
either complete or partial complementarity, are useful for the
design of new therapies against cancer polymorphism and viral
mutation. Currently over 1000 native miRNA species found in
vertebrates and many more new miRNA homologs continue to be
identified; however, most of their functions remain to be
determined. In this book, many new perspectives of the miRNA
research are reviewed and discussed, including their roles in stem
cell maintenance, embryonic development, tissue differentiation,
adult physiology, disease pathology, cancer research, viral
infection, genetic engineering in plants, and utility in cosmetic
applications. These new findings may not only provide significant
insight into the various mechanisms of miRNAs but also offer a
great opportunity in developing new therapeutic interventions.
MicroRNA Protocols provides diverse, novel, and useful descriptions
of miRNAs in several species, including plants, worms, flies, fish,
chicks, mice, and humans. These include some useful adaptations and
applications that could be relevant to the wider research community
who are already familiar with the identification of miRNAs. This
volume will stimulate the reader to explore diverse ways to
understanding the mechanism in which miRNAs facilitate the
molecular aspects of the biomedical research.
Since its invention and subsequent development nearly 20 years ago,
po- merase chain reaction (PCR) has been extensively utilized to
identify numerous gene probes in vitro and in vivo. However,
attempts to generate complete and full-length complementary cDNA
libraries were, for the most part, fruitless and remained elusive
until the last decade, when simple and rapid methods were
developed. With current decoding and potential application of human
genome information to genechips, there are urgent needs for
identification of functional significance of these decoded gene
sequences. Inherent in bringing these app- cations to fruition is
the need to generate a complete and full-length cDNA library for
potential functional assays of specific gene sequences. Generation
of cDNA Libraries: Methods and Protocols serves as a laboratory
manual on the evolution of generation of cDNA libraries, covering
both ba- ground information and step-by-step practical laboratory
recipes for which p- tocols, reagents, operational tips,
instrumentation, and other requirements are detailed. The first
chapter of the book is an overview of the basics of generating cDNA
libraries, which include the following: (a) the definition of a
cDNA library, (b) different kinds of cDNA libraries, (c)
differences between methods for cDNA library generation using
conventional approaches and novel stra- gies, including reverse
generation of RNA repertoires from cDNA libraries, and (d) the
quality of cDNA libraries.
This volume focuses on the analysis of miRNA and its many
components such as targets and expression profiling, regulation of
gene expression, miRNA detection in biofluids, and its application
in cancer and wound healing. The chapters in this book explore
topics that's cover biogenesis, isolation and profiling of miRNA,
exosomal miRNAs, and the study of the functional significance of
miRNAs. The chapters also cover screening miRNAs in mR-302/367
induced iPSCs, exosomal miRNAs from iPSC-derived cardiomyocytes
proliferation, and the recently discovered sugar-like RNA
glycylglycerins. Written in the highly successful Methods in
Molecular Biology series format, chapters include introductions to
their respective topics, lists of the necessary materials and
reagents, step-by-step, readily reproducible laboratory protocols,
and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, MicroRNA Protocols, Third Edition is a
valuable resource that provides researchers with a comprehensive
study of miRNAs and its numerous components and uses.
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