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本帖最后由 细胞海洋 于 2010-9-19 20:18 编辑
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PCR 应该是分子生物学最基本的实验手段，但并不是每个实验都有一帆风顺的，基于PCR的实验越来越多，SSCP，定点突变，
+ [/ ?4 L* R! \等 PCR进行基因沉默的制备。比较实用的基因手段。大家根据需要选择下载，希望有所帮忙。
- K! z' X" T6 f! _5 |Preface .............................................................................................................v
- c0 Y6 {/ |/ Z$ m6 N( @; uContributors .....................................................................................................xi
# W# _9 T2 H: ]" }. aPART I. PERFORMING AND OPTIMIZING PCR
% [ F# m% l) F9 N3 U) \9 N1 Polymerase Chain Reaction: Basic Principles and Routine Practice
4 D% W# u# u6 c6 G0 F3 M! s9 [Lori A. Kolmodin and David E. Birch .................................................. 3/ ~) o' ]& z1 ~: Z6 N
2 Computer Programs for PCR Primer Design and Analysis
: B8 p$ W* |+ t, x/ G/ wBing-Yuan Chen, Harry W. Janes, and Steve Chen ........................ 19
: z; ~" j# ^ Q* x3 Single-Step PCR Optimization
1 q, V5 D, t+ QUsing Touchdown and Stepdown PCR Programming0 f! D% _ Y3 a: L
Kenneth H. Roux .................................................................................. 31
5 J3 I8 @5 Z1 [4 XL PCR Amplification of Long Targets from Genomic DNA
8 o7 ]' Y8 v- ]Lori A. Kolmodin .................................................................................. 37, w5 T% ~4 t+ Y3 F
5 Coupled One-Step Reverse Transcription and Polymerase Chain
! k* @8 I% W. D; c( b+ ]( F6 WReaction Procedure for Cloning Large cDNA Fragments4 l0 E- b- q/ [/ t
Jyrki T. Aatsinki ................................................................................... 53
' i( |3 w) a9 V8 b6 Long Distance Reverse-Transcription PCR, q: M1 l. C2 A( x, y- B
Volker Thiel, Jens Herold, and Stuart G. Siddell ............................. 59
! `6 J7 S x0 R9 [* Q7 Increasing PCR Sensitivity for Amplification
5 C& H; L( C! J9 ?9 G6 f' Ufrom Paraffin-Embedded Tissues) X; e C' T; ~
Abebe Akalu and Juergen K. V. Reichardt ....................................... 67" E9 K9 ^. n$ a7 x$ P b; C, a
8 GC-Rich Template Amplification by Inverse PCR:
% Y) L- l( q1 w3 e, VDNA Polymerase and Solvent Effects
/ r5 c9 c6 } z) P4 UAlain Moreau, Da Shen Wang, Steve Forget, Colette Duez," ~ s7 Y2 C. k" s2 o
and Jean Dusart............................................................................... 75
8 _. F" d6 K0 K# h* L9 PCR Procedure for the Isolation of Trinucleotide Repeats
3 d# J0 X. E& I' fTeruaki Tozaki ...................................................................................... 81
8 U( n) v; r) h* q! L @( J10 Methylation-Specific PCR
$ a* L$ r2 R+ v$ \Haruhiko Ohashi .................................................................................. 912 z9 \! m# X$ [% S7 u
11 Direct Cloning of Full-Length Cell Differentially Expressed Genes
" S+ [8 v7 m9 z# @+ kby Multiple Rounds of Subtractive Hybridization
. S6 i% `* r+ E- q* R( ~) G( J' g) \Based on Long-Distance PCR and Magnetic Beads
! g9 K8 h3 e8 Q$ ^0 yXin Huang, Zhenglong Yuan, and Xuetao Cao ................................ 998 t c& Z( S% w$ Q9 X% x
PART II. CLONING PCR PRODUCTS: O5 b% p+ u% b: r$ ^
12 Cloning PCR Products: An Overview
/ j; G' A8 j5 f/ u" H8 Y5 pBaotai Guo and Yuping Bi ................................................................ 111
0 Z$ D1 N+ T; G0 U* V13 Using T4 DNA Polymerase to Generate Clonable PCR Products
& c. w, h( a' \. ^ a$ XKai Wang ............................................................................................. 121
0 `! K8 F# f2 s1 k' j5 c" W. G14 Enzyme-Free Cloning of PCR Products
1 Y2 h9 ]9 j3 c6 @and Fusion Protein Expression
. @$ G: Z2 h7 H) l3 T7 I# `1 P' vBrett A. Neilan and Daniel Tillett ..................................................... 125
; M7 x1 e) @' p7 {! A) B' ]6 d15 Directional Restriction Site-Free Insertion of PCR Products3 c: c! n' K0 j! _, n& T
into Vectors0 g. A) N) \6 Q
Guo Jun Chen .................................................................................... 133" d( [8 V' j5 v( I1 p
16 Autosticky PCR:
( J2 K3 g3 p6 E: i6 |3 Q/ @0 `6 eDirectional Cloning of PCR Products with Preformed 5' Overhangs3 ~5 V; T; V4 z
József Gál and Miklós Kálmán......................................................... 141
. c$ ? g/ h; B: {17 A Rapid and Simple Procedure for Direct Cloning4 b+ j6 T% p3 [2 `
of PCR Products into Baculoviruses( n+ U. q, @( L' N) a9 J) H; @
Tamara S. Gritsun, Michael V. Mikhailov,% ~# F9 x( Z6 z% c
and Ernest A. Gould ...................................................................... 153
% N: ?3 Z) b2 t* Y7 JPART III. MUTAGENESIS AND RECOMBINATION
+ y' W$ n0 g2 L, U0 K5 n18 PCR Approaches to DNA Mutagenesis and Recombination:
3 i4 @* A* L8 m% b& qAn Overview
5 l4 U- n0 D6 s, p0 mBinzhang Shen ................................................................................... 1678 |2 i3 n2 ~: T% {
19 In-Frame Cloning of Synthetic Genes Using PCR Inserts
: Y: `% f: w3 L2 m+ V3 }. SJames C. Pierce ................................................................................. 175
$ g* J# t/ K# Z5 j9 n7 z20 Megaprimer PCR% a; O* K T% |/ |; Z" \
Sailen Barik ........................................................................................ 189
& [) d3 D- N3 `8 y2 f0 O21 PCR-Mediated Recombination:
, c+ K- [& d2 Y7 ~A General Method Applied to Construct Chimeric Infectious
$ l4 F% ]- }0 S# MMolecular Clones- m2 `% t% R9 k( J3 |$ Y$ T# {
Guowei Fang, Barbara Weiser, Aloise Visosky, Timothy Moran,3 h+ K$ C4 L% n! W9 I' n2 p5 J
and Harold Burger ......................................................................... 1970 ^( i* x! G% Z8 L
22 PCR Method for Generating Multiple Mutations at Adjacent Sites
5 v5 f2 g& p4 |$ lJiri Adamec ......................................................................................... 2073 z% [& p8 j+ j& C
23 A Fast Polymerase Chain Reaction-Mediated Strategy for Introducing
- |0 f5 l* N8 n+ O+ d: _/ TRepeat Expansions into CAG-Repeat Containing Genes v% n2 q7 W, d" x" K2 w1 \# v
Franco Laccone ................................................................................. 217
4 f( S4 R+ h5 \0 _1 F, p24 PCR Screening in Signature-Tagged Mutagenesis of Essential Genes
" }# l: a9 b: W, xDario E. Lehoux and Roger C. Levesque ....................................... 225
: o0 |6 V0 {% w" X0 s6 k' Q25 Staggered Extension Process (StEP) In Vitro Recombination
* \/ u+ A8 D' `0 f' lAnna Marie Aguinaldo and Frances Arnold ................................... 2357 C3 r; x( M6 A7 {( E. r
26 Random Mutagenesis by Whole-Plasmid PCR Amplification
8 } b+ b- A$ D* KDonghak Kim and F. Peter Guengerich .......................................... 241" i! y* u& f& Y6 w7 V
PART IV. CLONING UNKNOWN NEIGHBORING DNA/ e/ H' F+ q- E; ] p$ Y
27 PCR-Based Strategies to Clone Unknown DNA Regions
+ @7 X1 n' {4 a& t" sfrom Known Foreign Integrants: An Overview
5 W* Z+ z+ {) b0 CEric Ka-Wai Hui, Po-Ching Wang, and Szecheng J. Lo ................ 2496 q) ]5 u$ Q' R" f9 O3 l
28 Long Distance Vectorette PCR (LDV PCR) h9 H. r6 Z8 H: `* K4 q# w
James A. L. Fenton, Guy Pratt, and Gareth J. Morgan ................. 275% J: N7 Q( _; V/ b6 H: w
29 Nonspecific, Nested Suppression PCR Method& e5 W. H8 ]3 p& z& I
for Isolation of Unknown Flanking DNA (“Cold-Start Method”)& J3 H' ^0 Q) g. B/ C" ~# l2 l
Michael Lardelli .................................................................................. 285
; A: f$ L! F( i$ M+ I2 g30 Inverse PCR: cDNA Cloning
7 X+ i, Z( a. u: f" OSheng-He Huang ................................................................................ 293
* p0 k% F& U' e8 X4 W8 {31 Inverse PCR: Genomic DNA Cloning
0 I* H2 [) e( d! YAmbrose Y. Jong, Anna T’ang, De-Pei Liu,
. w5 E' @, y# Dand Sheng-He Huang .................................................................... 301! N- c: B) j' ?0 R+ n+ K& O
32 Gene Cloning and Expression Profiling by Rapid Amplification
9 w* \% u6 S, ~6 q! Y) xof Gene Inserts with Universal Vector Primers
8 m( W; P; z+ G# e" qSheng-He Huang, Hua-Yang Wu, and Ambrose Y. Jong .............. 309+ I$ f$ e; g _8 v! {$ p& n5 y
33 The Isolation of DNA Sequences Flanking Tn5 Transposon Insertions5 B; i" {% `* A/ u8 B, f
by Inverse PCR6 i- l. h/ C1 L
Vincent J. J. Martin and William W. Mohn ...................................... 315& [; u" x5 K# T* h
34 Rapid Amplification of Genomic DNA Sequences Tagged4 E8 R3 z2 y( |0 N! s& ?
by Insertional Mutagenesis
) s2 w7 q( K) b5 G1 z' v ]Martina Celerin and Kristin T. Chun ................................................ 325+ N2 H5 \2 R4 C* O/ m
35 Isolation of Large Terminal Sequences of BAC Inserts Based
9 e+ Q: I- S/ S% ton Double-Restriction-Enzyme Digestion Followed# s w- o4 U+ ?6 c
by Anchored PCR1 W1 @* ^5 F; W* v, F1 W
Zhong-Nan Yang and T. Erik Mirkov ............................................... 337 {# E: V) D# G' h9 N5 F, j4 ^
36 A “Step Down” PCR-Based Technique for Walking) C {7 O& r" c& h4 W0 d8 K
Into and the Subsequent Direct Sequence Analysis2 a N ^% e$ }# `
of Flanking Genomic DNA
2 o* |! x% z; Y; z9 r5 CZiguo Zhang and Sarah Jane Gurr .................................................. 343
! j- v9 q2 n' v7 `. l# W6 OPART V. LIBRARY CONSTRUCTION AND SCREENING& n9 E e" V% }( }) ^* Y' x
37 Use of PCR in Library Screening: An Overview
4 A. m$ H# D& M6 p/ MJinbao Zhu .......................................................................................... 353$ H% U+ C* t1 D
38 Cloning of Homologous Genes by Gene-Capture PCR
" z1 k' w/ M- ^Renato Mastrangeli and Silvia Donini ............................................. 359
* z5 R; S- z8 h39 Rapid and Nonradioactive Screening of Recombinant Libraries by PCR0 f! P: C3 X* d8 ]" D
Michael W. King ................................................................................. 377
$ b* U! v& E# g& k3 u40 Rapid cDNA Cloning by PCR Screening (RC-PCR)
. E8 S* V6 S( R& H& U W5 M) f7 |3 EToru Takumi ....................................................................................... 3858 ?; z, P7 w$ ]; `* B8 T& \
41 Generation and PCR Screening of Bacteriophage λ Sublibraries2 B, A% L% E/ z+ x4 e" b8 h
Enriched for Rare Clones (the “Sublibrary Method”)
0 O6 c: J# n) W* x; ?2 EMichael Lardelli .................................................................................. 391 {! L6 P, D$ }) ^- E
42 PCR-Based Screening for Bacterial Artificial Chromosome Libraries
2 l r, S3 C/ z- C1 A y! ?- c: {Yuji Yasukochi ................................................................................... 401
9 x6 ?) G. X9 V, m2 }4 R43 A 384-Well Microtiter-Plate-Based Template Preparation' _8 t7 ~+ \4 S" l; R; |
and Sequencing Method
4 w7 |( ?* x$ m8 `" g& GLei He and Kai Wang ......................................................................... 411
$ U* ^, k1 t* c3 \3 Y44 A Microtiter-Plate-Based High Throughput PCR Product, U( u/ X: U9 c, D
Purification Method$ l& F! _4 ~ b, ^% [+ Q
Ryan Smith and Kai Wang ................................................................ 417
* `. Z' C; c( W r# ^Index ............................................................................................................ 423
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发表于 2010-9-19 18:24
