. j+ c( O. F1 r) V( q+ [6 G, Y. \4 dPreface: S' d( c" d k7 G2 m8 s8 }; k
Thirty years ago, the investigation of gene sequences and, in particular, disease-causing! `% K* a0 H+ u
mutations was a tedious, highly skilled operation; however, this field was revolutionized in 9 G/ ~# D3 R6 v, ^the mid-1980s with the introduction and subsequent development of a technique which9 Y' U d; ?( ~ U
enabled the reliable amplification of minute quantities of starting DNA into readily detectable6 {- }! }, S3 n2 D
levels. Nowadays, this technique, otherwise known as reverse-transcription polymerase9 o0 F" D7 E, q& c
chain reaction (RT-PCR), has become routine in most laboratories. Indeed, such / g; @, s7 P( R: h0 X4 T/ }is the popularity of RT-PCR that a protocol has been published describing how this could) s& X7 x2 C6 N9 e8 K5 W3 l
be carried out in a kitchen using common household equipment. The aim of this volume : Z& n2 u. O( h1 y/ Jis to translate RT-PCR theory into practice. To achieve this, a comprehensive guide to b. r, y! @& I. Fcurrently available RT-PCR techniques is given in the form of user-friendly protocols. 4 z5 V+ w! q" ^1 K: z3 ^6 s4 ]These protocols contain precise information about all the necessary chemical, consumable, 0 n+ j9 t4 y8 \$ P+ wand equipment resources and detailed instructions about how to perform each stage 8 K, A. O- b. e+ k( Hof the different methods. Furthermore, each protocol concludes with a comprehensive 8 r$ h; b& L, q0 z" o' Vnotes section, where authors provide helpful hints, trouble-shooting tips, and other mustknow( W# G; }7 Y+ g( y1 R& i
information in a format which is accessible to the beginner. 1 M8 h( a3 Q9 c7 [# jRT-PCR protocols were the subject of an earlier edition in the Methods in Molecular " ?, `8 k- L6 D u# @! K; [' oBiology™ series, which was published in August 2002. In this second edition, some of the6 @! s- H$ i; w
contents of the previous edition are revisited bringing these technologies up to date, for : X! l2 `2 V1 k! ~) A9 j5 K }( mexample, competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR $ U. l3 n. [8 ufor cloning. In addition, the second volume also describes the newer technologies that have+ A' e4 G: s) c L! @) [3 e
been developed and applied in the last 7 years including multiplex RT-PCR and RT-LATEPCR.% k4 O, x" G4 I; R# u( B; v% ?
This growth and development is reflected in the wide selection of basic RT-PCR 3 d+ h; b3 N; Utechniques which are presented in the first section entitled “The RT-PCR Detective: Hunting " e4 G6 B0 T( T. _Down the Right Method”. Arguably, however, the greatest advances in RT-PCR have come 7 u0 m6 J p, Sin the field of real-time quantitative RT-PCR, and this, along with all the other means of ( x% |2 ~3 n2 ~( L( `5 j. Vquantifying PCR products, is explained in the second section, “The RT-PCR Mathematician:) h$ h) o) U1 D& W5 u' V0 L
Assessing Gene and RNA Expression”. Finally, since designing RT-PCR experiments requires 2 Q+ \# Y$ H4 c* W M) |9 s8 O0 Wboth the correct recipe and the best ingredients, the last section, “The RT-PCR Master 0 V n ^8 B2 }! x& d7 VChef: Finding the Best Ingredients,” is devoted to recent advances in some of the individual . L% b4 y9 Z+ U" r3 `elements that go together to make the optimum RT-PCR reaction, e.g. RNA extraction,. |! |% F% K+ U5 h" a
primer design, and reverse transcription.0 _0 h) x# I8 e% B2 O! X5 q1 p* n
This volume is not intended to be a hard-core technical manual that is accessible to a# |' u! @$ {8 B( K
few Nobel Laureate molecular biologists. Rather, the goal is that it should act as a handy Y- K% w: y! |7 M' F
companion to anyone who wants to explore the marvels of gene expression. This includes4 [2 E Z; @' Z& Q1 O {
students and their tutors, researchers, laboratory managers, and technologists from many3 X3 Q4 f6 c- g! Y. P4 _
diverse disciplines ranging from biochemistry to zoology and forensics to physiology.6 R- L% i& X* {. a2 C" c d9 d( b
y! h6 p" b9 q3 V8 Y
Armidale, NSW : l7 G( e$ Q$ B: ]2 a4 p4 f
Nicola King5 U' \4 U# M+ X2 ?+ |5 s7 H, c9 v
, C! N4 S; u/ |$ [9 x4 E. ]0 P1 F 作者: uyunbilig 时间: 2010-8-6 15:24
