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本帖最后由 细胞海洋 于 2010-9-19 20:18 编辑 5 X: U5 c* w: X- D' ~
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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，5 |( {4 Q4 A: }
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。
1 e) I# o7 i2 i5 uPreface .............................................................................................................v
, M: `( P3 b) nContributors .....................................................................................................xi
m+ c Z }% ?/ ~) aPART I. PERFORMING AND OPTIMIZING PCR
$ C" Z/ t4 W# @7 R1 Polymerase Chain Reaction: Basic Principles and Routine Practice, I* {) a o8 W0 m7 H
Lori A. Kolmodin and David E. Birch .................................................. 3
$ Z+ m2 v! _. X, x5 ~- N2 Computer Programs for PCR Primer Design and Analysis# J) N+ j' r' {: P6 G7 g# j# t
Bing-Yuan Chen, Harry W. Janes, and Steve Chen ........................ 19
3 n9 O, C, W1 w" @3 Single-Step PCR Optimization
, o7 H# p' ^9 R8 v, X8 v: UUsing Touchdown and Stepdown PCR Programming
4 E5 E# ]; B* S: AKenneth H. Roux .................................................................................. 31
5 R0 S4 [3 J; R- X4 XL PCR Amplification of Long Targets from Genomic DNA; m" b3 E6 G. x* p% q2 T' \6 w# N
Lori A. Kolmodin .................................................................................. 37+ Y! `6 G! y# U' x6 c, c# m# j
5 Coupled One-Step Reverse Transcription and Polymerase Chain2 Q. v/ Y/ Q5 v( X' s9 L' Z
Reaction Procedure for Cloning Large cDNA Fragments+ H" c# K/ ~8 Z7 y: Y; {; E
Jyrki T. Aatsinki ................................................................................... 53
2 m- w! O J5 W. h* N6 Long Distance Reverse-Transcription PCR
: S- x; J1 q/ i1 i8 iVolker Thiel, Jens Herold, and Stuart G. Siddell ............................. 59
% R. ~7 {( Q* M3 ]! Y( A7 Increasing PCR Sensitivity for Amplification
* h `3 I3 `* {7 K$ Nfrom Paraffin-Embedded Tissues
, D6 k! e; W; w) G5 jAbebe Akalu and Juergen K. V. Reichardt ....................................... 67) u5 D* r) w* A+ h1 F# p3 @
8 GC-Rich Template Amplification by Inverse PCR:2 {( _) P: c. t6 O' a8 M3 p& e
DNA Polymerase and Solvent Effects
% O0 d! @: f: @# d hAlain Moreau, Da Shen Wang, Steve Forget, Colette Duez,
$ y$ |3 Z7 U2 @3 Y* d; X/ mand Jean Dusart............................................................................... 75
7 t, Y2 l4 g7 ~& [. C/ O4 @" ?7 r9 PCR Procedure for the Isolation of Trinucleotide Repeats, k; u# e' U2 J4 R1 e# ~
Teruaki Tozaki ...................................................................................... 814 h: z1 x/ d( y I) N8 I- {
10 Methylation-Specific PCR' a V ~! \+ t1 w
Haruhiko Ohashi .................................................................................. 91
( A0 K" u" p6 D" c3 B; P11 Direct Cloning of Full-Length Cell Differentially Expressed Genes
3 T/ s# P% _& W( V7 gby Multiple Rounds of Subtractive Hybridization5 G4 Q1 i7 w0 Q( z9 R. P( T
Based on Long-Distance PCR and Magnetic Beads
0 i" ^! U" I _. f7 q! OXin Huang, Zhenglong Yuan, and Xuetao Cao ................................ 99
% t2 e7 w8 b: y% H. zPART II. CLONING PCR PRODUCTS. ?$ x# G, B! I6 q2 }3 _9 v# A
12 Cloning PCR Products: An Overview
% K- Y# N2 P2 O. O- n% ]Baotai Guo and Yuping Bi ................................................................ 1112 J$ f q- \! R' b# |7 g/ Y9 {
13 Using T4 DNA Polymerase to Generate Clonable PCR Products
, ]5 [' @8 @- }* {5 h) n7 |& nKai Wang ............................................................................................. 121. s4 y3 U# N$ P! F
14 Enzyme-Free Cloning of PCR Products
+ ^- K% j/ j/ @# j) i# f8 Mand Fusion Protein Expression
+ k: z' @9 |+ R4 f; lBrett A. Neilan and Daniel Tillett ..................................................... 1253 i% s I0 z/ W5 Q+ Z1 p
15 Directional Restriction Site-Free Insertion of PCR Products* m: O* s8 z" _: X' y4 I( K7 J
into Vectors
. s/ |' g6 o$ _) TGuo Jun Chen .................................................................................... 133
/ y# Z* ]4 v- j' F, G" q1 b# x16 Autosticky PCR:
6 w3 L. B& G0 V5 _ m( o) J0 UDirectional Cloning of PCR Products with Preformed 5' Overhangs) N1 _5 B5 r. Z- B/ @2 v& K- k
József Gál and Miklós Kálmán......................................................... 1411 `( G) ~% p/ o* o
17 A Rapid and Simple Procedure for Direct Cloning3 t4 S* C7 w- k* N+ t* n8 h
of PCR Products into Baculoviruses
8 M( X/ e( [* h# [, K* }: tTamara S. Gritsun, Michael V. Mikhailov,/ @4 M2 Q% ~9 z. X$ n( h" V
and Ernest A. Gould ...................................................................... 153+ x; k9 g1 P+ j! y
PART III. MUTAGENESIS AND RECOMBINATION2 |4 E* B4 x% q
18 PCR Approaches to DNA Mutagenesis and Recombination:- A U* x3 d q9 t s5 ~
An Overview5 F$ L# A6 c' D/ y9 q0 x3 B
Binzhang Shen ................................................................................... 167
1 b. _. n+ \ n+ h* b19 In-Frame Cloning of Synthetic Genes Using PCR Inserts
3 R% g+ Q% J. u# M, U# k2 k* `- ?James C. Pierce ................................................................................. 175. e9 O3 f, p8 X7 m5 d+ I
20 Megaprimer PCR
- m% Q/ \8 {7 p* }* V4 |! G# T& NSailen Barik ........................................................................................ 189/ Y+ S8 A& I. L' K! W% u; _6 u
21 PCR-Mediated Recombination:
* Q; h8 X0 \" F, F# lA General Method Applied to Construct Chimeric Infectious1 G' W) G! n+ O ^% ^1 R
Molecular Clones
% K6 _6 G. ~! W6 b( [Guowei Fang, Barbara Weiser, Aloise Visosky, Timothy Moran,
- `$ x$ {/ z$ _( J% T. \$ dand Harold Burger ......................................................................... 197
2 I+ \2 v. j* e. \22 PCR Method for Generating Multiple Mutations at Adjacent Sites6 k: {6 z1 }$ z4 @1 [, M
Jiri Adamec ......................................................................................... 207* n2 n, _& }$ g. R+ V/ c, g
23 A Fast Polymerase Chain Reaction-Mediated Strategy for Introducing( m) Y6 U6 R9 n3 V; k
Repeat Expansions into CAG-Repeat Containing Genes% E9 C3 q, ~) q& F$ i0 B
Franco Laccone ................................................................................. 217# ?2 t. ~/ A) ~$ V- x
24 PCR Screening in Signature-Tagged Mutagenesis of Essential Genes9 ], ~0 _: v) C3 t3 I! a1 K
Dario E. Lehoux and Roger C. Levesque ....................................... 2250 }! {# F+ E1 u9 H4 ?3 a
25 Staggered Extension Process (StEP) In Vitro Recombination
) o4 `% T% R" Q+ P1 ]# G# E5 }! uAnna Marie Aguinaldo and Frances Arnold ................................... 235" V. E# H) B7 D7 W; l* p
26 Random Mutagenesis by Whole-Plasmid PCR Amplification
" d$ ]) d; J; s0 m( R, @! B1 }; RDonghak Kim and F. Peter Guengerich .......................................... 241
: H! V" Y$ J1 I/ S- |" yPART IV. CLONING UNKNOWN NEIGHBORING DNA: f5 Y4 h+ ?* E" |1 ~0 ^" L
27 PCR-Based Strategies to Clone Unknown DNA Regions
( k! T1 a$ i$ A' d+ ifrom Known Foreign Integrants: An Overview6 V9 d m$ J- b6 R8 E9 X9 ?- i
Eric Ka-Wai Hui, Po-Ching Wang, and Szecheng J. Lo ................ 2497 K( b+ w. s, K( f/ K, p
28 Long Distance Vectorette PCR (LDV PCR)
; a& L* s0 q8 V# e# wJames A. L. Fenton, Guy Pratt, and Gareth J. Morgan ................. 2758 l# S/ f# J: z2 B8 P
29 Nonspecific, Nested Suppression PCR Method+ I5 z$ r3 K& F# A4 R
for Isolation of Unknown Flanking DNA (“Cold-Start Method”)8 e& L% s- Z5 W9 [ @4 p C* \9 \
Michael Lardelli .................................................................................. 285
2 F% v# h, O; \3 B- z30 Inverse PCR: cDNA Cloning6 t# N5 y q: ?& u! b2 E1 g' ~
Sheng-He Huang ................................................................................ 293' Q! }) C; d$ `( X7 K) d
31 Inverse PCR: Genomic DNA Cloning
8 ?: C) H9 W" w* @; A$ ^2 [Ambrose Y. Jong, Anna T’ang, De-Pei Liu,
; h# r0 z" v8 t+ Dand Sheng-He Huang .................................................................... 301
# @" O9 Q/ r( H32 Gene Cloning and Expression Profiling by Rapid Amplification/ F9 ^* t6 O' Y; [
of Gene Inserts with Universal Vector Primers
$ {2 w+ J+ R( c- D" H! P; m% rSheng-He Huang, Hua-Yang Wu, and Ambrose Y. Jong .............. 309
; k# Z/ N1 i3 Z! D. i) B) n33 The Isolation of DNA Sequences Flanking Tn5 Transposon Insertions
7 e3 {- ?3 i. }( W) x" n4 r2 h# k$ jby Inverse PCR
( _* U* n# _. T9 D' f$ T4 c! dVincent J. J. Martin and William W. Mohn ...................................... 315/ g# d" ~1 D) T; d( ?
34 Rapid Amplification of Genomic DNA Sequences Tagged: I- q8 o& s. a0 @) y
by Insertional Mutagenesis3 \ j/ _, F2 F0 T4 V D
Martina Celerin and Kristin T. Chun ................................................ 325
) s5 v3 U6 J; q" a; t35 Isolation of Large Terminal Sequences of BAC Inserts Based
1 |8 M! t8 A2 ^% D non Double-Restriction-Enzyme Digestion Followed% ]3 {6 l% E7 ]8 a
by Anchored PCR+ D# j, n8 w/ a
Zhong-Nan Yang and T. Erik Mirkov ............................................... 337
) P* {' O# I C$ P4 n/ {; [$ b0 b36 A “Step Down” PCR-Based Technique for Walking
, |! y2 T+ s/ w7 P. BInto and the Subsequent Direct Sequence Analysis
/ w' X' p/ I& q: J! R* Q) Wof Flanking Genomic DNA0 _% A$ W V4 C8 u1 @3 C8 S9 t
Ziguo Zhang and Sarah Jane Gurr .................................................. 343
6 K& L( A2 h6 }2 `PART V. LIBRARY CONSTRUCTION AND SCREENING
& R8 s f) c1 ~) f; _, f37 Use of PCR in Library Screening: An Overview) F9 [1 O* {; j; h9 }/ s
Jinbao Zhu .......................................................................................... 3535 f9 @# s+ @& i" }0 |/ T
38 Cloning of Homologous Genes by Gene-Capture PCR1 i, ~& x7 y/ y+ l/ n7 ?
Renato Mastrangeli and Silvia Donini ............................................. 359
' Q% P9 K+ t; Q8 K4 |0 U# @39 Rapid and Nonradioactive Screening of Recombinant Libraries by PCR
* p8 Q+ z/ l7 {8 R, A; E: Z+ EMichael W. King ................................................................................. 3776 S& \. c l7 Q+ \+ U* O- J
40 Rapid cDNA Cloning by PCR Screening (RC-PCR). }0 {( F+ x# Q% B( M* P
Toru Takumi ....................................................................................... 385
( ?3 j8 j+ n( u0 `& \. l41 Generation and PCR Screening of Bacteriophage λ Sublibraries
# K% u! G ?" ~1 EEnriched for Rare Clones (the “Sublibrary Method”)& F q( x& @% e d; x
Michael Lardelli .................................................................................. 391
6 N' R+ \7 c' b: \2 h, q0 w42 PCR-Based Screening for Bacterial Artificial Chromosome Libraries
& w" p, V/ a( `* bYuji Yasukochi ................................................................................... 401
8 c e; ^$ S" `43 A 384-Well Microtiter-Plate-Based Template Preparation: H2 _6 H, _9 k( j7 o f
and Sequencing Method
; [, f& K( W/ ]) ^7 ] ^* O3 E, cLei He and Kai Wang ......................................................................... 411 p* _/ u3 @0 o: }' [# B
44 A Microtiter-Plate-Based High Throughput PCR Product
( q! S' g* e. a$ wPurification Method
8 p! |5 [- i' c( q5 ]) |" {0 a* lRyan Smith and Kai Wang ................................................................ 417' m5 w- W6 h' `/ l/ L
Index ............................................................................................................ 423+ c7 \* o4 O3 {
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