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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RT-PCR Protocols7 L; b& s: ]$ w6 C* h9 e7 s
Second Edition
9 G9 s, K2 E' A. G, g3 E. ]8 S' xEdited by Nicola King
) s3 W7 l( w: H1 |& o' KUniversity of New England, School of Science and Technology, Armidale, Australia
, `; _8 s/ v4 e8 o6 N5 ]ISSN 1064-3745 e-ISSN 1940-6029% O" L1 B1 j! x3 E
ISBN 978-1-60761-628-3 e-ISBN 978-1-60761-629-0/ t8 D3 c( h1 Y% x6 r4 o8 v0 y
DOI 10.1007/978-1-60761-629-0
& K: `( B0 ], {9 ~0 F$ p  ^6 XSpringer New York Dordrecht Heidelberg London& T8 C' q+ U: @- P9 |
Library of Congress Control Number: 20109235975 a/ X; h0 K7 H) a
© Springer Science+Business Media, LLC 2010. F+ S. B! r$ v4 S
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Preface3 }" A5 G/ q+ a8 B- _' b
Thirty years ago, the investigation of gene sequences and, in particular, disease-causing
6 a* ]4 r( l: `& gmutations was a tedious, highly skilled operation; however, this field was revolutionized in' Q& b0 U8 Z9 a/ p
the mid-1980s with the introduction and subsequent development of a technique which* V' u; p5 Q% M5 L) i. S
enabled the reliable amplification of minute quantities of starting DNA into readily detectable
; \) x3 {+ v) t: [levels. Nowadays, this technique, otherwise known as reverse-transcription polymerase) l$ K: j4 H5 f" o. a4 x* R' t: Z0 C: \- x
chain reaction (RT-PCR), has become routine in most laboratories. Indeed, such
; m- M/ w4 X2 s0 |6 _! v9 n* eis the popularity of RT-PCR that a protocol has been published describing how this could" J% M$ N% \4 v2 f& C
be carried out in a kitchen using common household equipment. The aim of this volume
0 i4 @  u0 h" D# n) s: \9 kis to translate RT-PCR theory into practice. To achieve this, a comprehensive guide to6 K/ ]" \! `' A$ `
currently available RT-PCR techniques is given in the form of user-friendly protocols.# M% m) }3 _2 g5 b+ a3 `! L9 w' t
These protocols contain precise information about all the necessary chemical, consumable,. |# l: @( W& ^
and equipment resources and detailed instructions about how to perform each stage1 e# ~* d% ?* X% t
of the different methods. Furthermore, each protocol concludes with a comprehensive
/ |0 h) N" {# v& ^# u$ A5 unotes section, where authors provide helpful hints, trouble-shooting tips, and other mustknow
7 }% Q2 Z2 P& m# Yinformation in a format which is accessible to the beginner.) s. K- Y2 a+ ?$ v  C( T
RT-PCR protocols were the subject of an earlier edition in the Methods in Molecular
8 p  J3 C" _9 j* K  {" z3 ZBiology™ series, which was published in August 2002. In this second edition, some of the1 |6 q9 P7 D/ G  O" C+ E
contents of the previous edition are revisited bringing these technologies up to date, for
: k; v: ?1 u' Y  h' R3 Yexample, competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR0 ]/ e8 W3 A. K5 p/ k& I( n
for cloning. In addition, the second volume also describes the newer technologies that have$ ?6 H; x7 a; V8 o8 K' j  V
been developed and applied in the last 7 years including multiplex RT-PCR and RT-LATEPCR.3 A+ B, [! K0 E' H4 }9 s
This growth and development is reflected in the wide selection of basic RT-PCR
- t* F# G7 J6 R5 f8 w  n, z. X) Btechniques which are presented in the first section entitled “The RT-PCR Detective: Hunting
3 v, r0 q+ G7 r) B) E6 [Down the Right Method”. Arguably, however, the greatest advances in RT-PCR have come& `7 `& Y& g% M! ~( z% G0 p9 I
in the field of real-time quantitative RT-PCR, and this, along with all the other means of, j% N. P- p4 S) I5 _9 m0 B& b
quantifying PCR products, is explained in the second section, “The RT-PCR Mathematician:
; p1 S3 A6 s7 y% a& ]( y% K  XAssessing Gene and RNA Expression”. Finally, since designing RT-PCR experiments requires
3 p- z9 i6 s3 f2 G1 T, zboth the correct recipe and the best ingredients, the last section, “The RT-PCR Master2 W. G. U6 u& u9 ^/ E2 K! |
Chef: Finding the Best Ingredients,” is devoted to recent advances in some of the individual0 p, s8 D. z" F2 {/ ?* I( c7 r
elements that go together to make the optimum RT-PCR reaction, e.g. RNA extraction,
% `% y( c" \. v, g- v% `- h8 h6 P: Oprimer design, and reverse transcription.
% N' a( O1 a' [# x! v: \This volume is not intended to be a hard-core technical manual that is accessible to a
2 d7 n4 Y  c, L  u" ^few Nobel Laureate molecular biologists. Rather, the goal is that it should act as a handy; R% _7 m/ `! d3 E; o
companion to anyone who wants to explore the marvels of gene expression. This includes( [9 }' j1 f" i2 @) a. z
students and their tutors, researchers, laboratory managers, and technologists from many; ~: U: D# S1 x0 P' T4 z
diverse disciplines ranging from biochemistry to zoology and forensics to physiology.
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Armidale, NSW + g3 Q. i4 s- b% J' J. Y5 r/ G# N
Nicola King% J/ W% O1 v" t
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