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Telomeres and Telomerase Methods and Protocols 1st
Edition Michael C. Bibby (Auth.) Digital Instant
Download
Author(s): Michael C. Bibby (auth.), John A. Double, Michael J. Thompson
(eds.)
ISBN(s): 9780896036574, 089603657X
Edition: 1st
File Details: PDF, 1.17 MB
Year: 2002
Language: english
Methods in Molecular Biology TM
VOLUME 191
Telomeres
and Telomerase
Methods and Protocols
Edited by
John A. Double
Michael J. Thompson
HUMANA PRESS
Telomeres and Telomerase
M E T H O D S I N M O L E C U L A R B I O L O G Y™
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M E T H O D S I N M O L E C U L A R B I O L O G Y™
Telomeres and
Telomerase
Methods and Protocols
Edited by
John A. Double
and
Michael J. Thompson
Cancer Research Unit, University of Bradford
Bradford, West Yorkshire, UK
Telomeres and telomerase : methods and protocols / edited by John A. Double and Michael J.
Thompson.
p. ; cm. -- (Methods in molecular biology ; v. 191)
Includes bibliographical references and index.
ISBN 0-89603-657-X (alk. paper)
1. Telomerase--Laboratory manuals. 2. Telomere--Laboratory manuals. I. Double, John A. II.
Thompson, Michael J.
[DNLM: 1. Telomere--physiology. 2. Telomerase--physiology. QH 600.3 T2777 2002]
QP606.T44 T45 2002
572.8'7--dc21
2001039598
Preface
v
Contents
Preface ................................................................................ v
Contributors ........................................................................ ix
1 Introduction to Telomeres and Telomerase
Michael C. Bibby .......................................................... 1
2 Detection of Chromosome Ends by Telomere FISH
Harry Scherthan ......................................................... 13
3 Telomere Length Distribution:
Digital Image Processing and Statistical Analysis
Jean-Patrick Pommier and Laure Sabatier ............. 33
4 Analysis of Telomerase RNA Gene Expression
by In Situ Hybridization
W. Nicol Keith, Joseph Sarvesvaran,
and Martin Downey ............................................... 65
5 Relative Gene Expression in Normal
and Tumor Tissue by Quantitative RT-PCR
Dennis S. Salonga, Kathleen D. Danenberg,
Jean Grem, Ji Min Park,
and Peter V. Danenberg ....................................... 83
6 Quantitative Detection of Telomerase Components
by Real-Time, Online RT-PCR Analysis
with the LightCycler
Thomas Emrich, Sheng-Yung Chang, Gerlinde Karl,
Birgit Panzinger, and Chris Santini.................... 99
7 Standard TRAP Assay
Angelika M. Burger .................................................. 109
8 Stretch PCR Assay
Jun-ichi Nakayama and Fuyuki Ishikawa ............. 125
vii
viii Contents
ix
x Contributors
1
Introduction to Telomeres and Telomerase
Michael C. Bibby
1. Cell Replication
Because of the mechanism of conventional DNA polymerases,
the replication of DNA molecules can be predicted to result in the
gradual shortening of the chromosome by the length of a terminal
primer at each cell cycle (1). This predication is supported by the
From: Methods in Molecular Biology, vol. 191: Telomeres and Telomerase: Methods and Protocols
Edited by: J. A. Double and M. J. Thompson © Humana Press Inc., Totowa, NJ
1
2 Bibby
2. Telomerase Structure
Telomerase is a specialized DNA polymerase that synthesizes
telomeric repeats de novo. It consists of an RNA subunit that acts as
the template for the synthesis of telomeric DNA, and this process is
catalyzed by a protein component (20). Therefore because telo-
merase polymerizes DNA it is a true reverse transcriptase. The RNA
component of telomerase was first characterized in ciliates (21, 22).
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