PDF电子书：RT-PCR Protocols 2nd ED 2010(MiMB 630)

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2010出版的 Methods in Molecular Biology 系列 vol.630
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  v% Y* W  u) ^- i+ aRT-PCR Protocols
1 R" z# f1 M( kSecond Edition
- b+ [6 B" J% V3 n! nEdited by Nicola King2 W% I. b1 A2 B2 c, t
University of New England, School of Science and Technology, Armidale, Australia! Z9 `5 {# M' I: b/ A
ISSN 1064-3745 e-ISSN 1940-6029% U: ^* \' y1 }5 [7 p
ISBN 978-1-60761-628-3 e-ISBN 978-1-60761-629-0
: ~- L, m" c% V( u7 b4 }. z( NDOI 10.1007/978-1-60761-629-04 I  A' H( t. V7 A) Y( G/ x+ M$ U
Springer New York Dordrecht Heidelberg London. ^+ J2 q4 }* l* G) c% ]- X. a* l, A% M
Library of Congress Control Number: 2010923597
* u- O4 w9 V. k  O© Springer Science+Business Media, LLC 2010
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Preface
4 P3 g# q  A/ y& l! TThirty years ago, the investigation of gene sequences and, in particular, disease-causing
2 g- N9 F9 ]6 `! s1 w9 ^- Ymutations was a tedious, highly skilled operation; however, this field was revolutionized in
* a- A* ~6 X: ~9 x) ]- Ethe mid-1980s with the introduction and subsequent development of a technique which, O, B, S! `2 c  M/ e) H
enabled the reliable amplification of minute quantities of starting DNA into readily detectable6 \$ A) Y/ l: {1 O/ E
levels. Nowadays, this technique, otherwise known as reverse-transcription polymerase
; o3 X% F/ ~# E$ \+ z0 ochain reaction (RT-PCR), has become routine in most laboratories. Indeed, such5 b6 }% P4 _! E0 C4 }, U
is the popularity of RT-PCR that a protocol has been published describing how this could
$ c& Y0 _9 B6 |, pbe carried out in a kitchen using common household equipment. The aim of this volume" d, t$ h3 a7 R+ b: w
is to translate RT-PCR theory into practice. To achieve this, a comprehensive guide to
* @6 C( n* u; E- J! \currently available RT-PCR techniques is given in the form of user-friendly protocols.% C# W1 Y9 z3 ^, y+ I3 ?
These protocols contain precise information about all the necessary chemical, consumable,! \  U, f0 o# y4 n! P
and equipment resources and detailed instructions about how to perform each stage
5 _, _* _  a- |5 [& v3 x3 Mof the different methods. Furthermore, each protocol concludes with a comprehensive6 `7 j% G. M& m' L7 Z
notes section, where authors provide helpful hints, trouble-shooting tips, and other mustknow: j- k9 d0 [5 a" T7 W8 u: q
information in a format which is accessible to the beginner.! d3 Q& z7 {1 Z7 E  ]4 X. i4 s0 b
RT-PCR protocols were the subject of an earlier edition in the Methods in Molecular7 @' o! w6 B6 ~* W% f
Biology™ series, which was published in August 2002. In this second edition, some of the2 @4 Z/ T: z) D* \
contents of the previous edition are revisited bringing these technologies up to date, for/ T: l! b' }5 v7 }( I
example, competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR2 H4 Y* q) v3 l5 Z3 u, `8 e
for cloning. In addition, the second volume also describes the newer technologies that have2 Y9 @  R9 C8 h" Y* _
been developed and applied in the last 7 years including multiplex RT-PCR and RT-LATEPCR.8 T8 j3 }% P+ a2 u
This growth and development is reflected in the wide selection of basic RT-PCR
/ J2 P. R+ u8 D4 K5 btechniques which are presented in the first section entitled “The RT-PCR Detective: Hunting5 c# X- T* P) S# S5 D2 d; o
Down the Right Method”. Arguably, however, the greatest advances in RT-PCR have come
/ L+ D+ b) q% @2 P: V6 }# x# Vin the field of real-time quantitative RT-PCR, and this, along with all the other means of6 `# @8 y) C3 c
quantifying PCR products, is explained in the second section, “The RT-PCR Mathematician:8 i; T) k" [7 X3 E% z3 a3 m1 g
Assessing Gene and RNA Expression”. Finally, since designing RT-PCR experiments requires; @, R* }$ O( R* N. L0 e  @2 X) z
both the correct recipe and the best ingredients, the last section, “The RT-PCR Master
6 K! }' H9 x" S. P; q5 g; kChef: Finding the Best Ingredients,” is devoted to recent advances in some of the individual
( F( P7 h/ S, [3 a+ delements that go together to make the optimum RT-PCR reaction, e.g. RNA extraction,: I- h6 O0 q0 X) @7 Y$ r5 ]
primer design, and reverse transcription.
6 z. U) N5 B4 ?1 X; ]: d% sThis volume is not intended to be a hard-core technical manual that is accessible to a
' x3 ~: e. f) Q5 y1 F, sfew Nobel Laureate molecular biologists. Rather, the goal is that it should act as a handy3 ~, Y! ?( d: f% S. s
companion to anyone who wants to explore the marvels of gene expression. This includes
9 G& E/ B" x; Q9 `% Z- Tstudents and their tutors, researchers, laboratory managers, and technologists from many# E# E" Q1 W! O  T) F
diverse disciplines ranging from biochemistry to zoology and forensics to physiology.* s0 e8 h1 P& m7 o) U

! x1 C1 V3 p& {0 ]/ X& x: g% oArmidale, NSW
9 W2 \, C) ~$ K3 b, R# P( P* C$ YNicola King
: ?" c9 E# `2 s2 x! o9 p* X! p, K( I0 j* _) q) c
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