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This volume includes comprehensive descriptions of miRNA biogenesis
and their role in the development and progression of various human
diseases. The first few chapters of MicroRNA Profiling: Methods and
Protocols discuss the effects of over-expressing and repressing of
a target miRNA and their effects on cell viability and
proliferation. The next few chapters explore the protocols for
total RNA isolation from cells and cell-derived product including
formalin fixed paraffin embedded tissue and plant tissue. The last
few chapters discuss isolation and characterization of exosomes
from medium conditioned by cell lines, serum, and plasma specimens.
This book also includes discussions of several software tools, such
as miRandola, PicTar, DIANA, and miRWalk. 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. Comprehensive and cutting-edge, MicroRNA
Profiling: Methods and Protocols is a valuable resource for anyone
interested in the field of Micro RNAs.
This volume includes comprehensive descriptions of miRNA biogenesis
and their role in the development and progression of various human
diseases. The first few chapters of MicroRNA Profiling: Methods and
Protocols discuss the effects of over-expressing and repressing of
a target miRNA and their effects on cell viability and
proliferation. The next few chapters explore the protocols for
total RNA isolation from cells and cell-derived product including
formalin fixed paraffin embedded tissue and plant tissue. The last
few chapters discuss isolation and characterization of exosomes
from medium conditioned by cell lines, serum, and plasma specimens.
This book also includes discussions of several software tools, such
as miRandola, PicTar, DIANA, and miRWalk. 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. Comprehensive and cutting-edge, MicroRNA
Profiling: Methods and Protocols is a valuable resource for anyone
interested in the field of Micro RNAs.
Diabetes is a chronic disorder of glucose metabolism and a major
cause of premature mortality. The potential use of replacement beta
cells as therapy for diabetes requires an ability to culture such
cells while maintaining their functional status. Glucose stimulated
insulin secretion (GSIS) is lost in long-term cultured MIN6
heterogeneous cells. MIN6 B1, a clonal sub-line derived from MIN6,
has been described as highly glucose-responsive. This study aimed
to investigate the GSIS function, changes in gene expression and,
subsequently, to develop possible experimental approaches to
overcome this loss. Understanding the molecular basis for loss of
GSIS may contribute to better culture conditions for islets in
transplantation programmes; it may also add to our understanding of
beta cell insensitivity to high blood glucose in Type 2 diabetes.
In parallel, in an attempt to identify more reliable biomarkers for
diabetes, we also investigated if extracellular mRNAs are
reproducibly detectable in conditioned medium (CM) from a range of
insulin-producing cell types and in serum specimens from Type 2
diabetes and controls.
This second edition provides updated and comprehensive methods on
miRNA biogenesis and their role in the development and progression
of various human diseases. Chapters detail miRNA biogenesis,
isolating RNA, extracellular vesicles (EVs), circulating miRNAs,
analyzing miRNA and miRDeep-P2, protocols for total RNA isolation
from cells, cell-derived products, isolation and characterization
of exosomes, serum, plasma specimens, and software tools. Written
in the 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 protocols, and notes on troubleshooting and avoiding
known pitfalls. Authoritative and cutting-edge, MicroRNA Profiling:
Methods and Protocols, Second Edition aims to provide comprehensive
and accessible methods to undergraduate, graduate, and established
scientist.
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