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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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0 H4 [% ~3 d. T- A d% PPCR 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，
3 Q6 K) \# l6 e5 ^; ` ?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。' a6 o' _4 d; Z7 m: N
Preface .............................................................................................................v
: ~$ q5 ~( s r* _! o, `' Z3 X$ N, |: EContributors .....................................................................................................xi7 x7 G3 j" e5 Z; L
PART I. PERFORMING AND OPTIMIZING PCR
; N S) ]3 {" H u/ A1 Polymerase Chain Reaction: Basic Principles and Routine Practice
! n/ F& P' j8 l6 }4 h1 M3 R3 G/ PLori A. Kolmodin and David E. Birch .................................................. 34 | i$ h& x' p# l
2 Computer Programs for PCR Primer Design and Analysis
5 Y. b; K+ x, MBing-Yuan Chen, Harry W. Janes, and Steve Chen ........................ 19
% g3 W5 K, E3 K& y B& ?3 Single-Step PCR Optimization
3 ~; U# b( s( H PUsing Touchdown and Stepdown PCR Programming
0 C+ z; j ]# Q1 j. VKenneth H. Roux .................................................................................. 31% l2 B; g }! a. M. u$ l# a
4 XL PCR Amplification of Long Targets from Genomic DNA
- {8 @1 n2 p- z& F) o) D$ L2 }Lori A. Kolmodin .................................................................................. 37" v9 t& ]; `6 ?+ g; g. _# L
5 Coupled One-Step Reverse Transcription and Polymerase Chain
4 N0 G$ I2 o; s5 tReaction Procedure for Cloning Large cDNA Fragments
- d; x9 F7 ? hJyrki T. Aatsinki ................................................................................... 535 B9 V! c. D* b
6 Long Distance Reverse-Transcription PCR
& a4 I: T* m/ [7 j9 |( I, tVolker Thiel, Jens Herold, and Stuart G. Siddell ............................. 59
$ C) a/ j( u; V g7 Increasing PCR Sensitivity for Amplification
& ~6 s$ X! ^. p% @from Paraffin-Embedded Tissues
: r+ ?4 b$ ~! G8 `( iAbebe Akalu and Juergen K. V. Reichardt ....................................... 67
. W; Y0 v6 q/ s+ K7 v" M8 GC-Rich Template Amplification by Inverse PCR:" t% A9 G2 I' b+ Z8 n; e' p# h6 y% X4 E
DNA Polymerase and Solvent Effects/ ^: G+ ?5 v2 I& f7 e8 h7 \
Alain Moreau, Da Shen Wang, Steve Forget, Colette Duez, C' _/ \) q5 ^5 h: \2 N- e2 C- }
and Jean Dusart............................................................................... 75: ?4 N* k+ S. N7 [; N
9 PCR Procedure for the Isolation of Trinucleotide Repeats
/ R7 r4 r# y* e6 u6 r+ _7 PTeruaki Tozaki ...................................................................................... 81
) {& x/ ?, f* k- T10 Methylation-Specific PCR
4 K- n. v) j: ~! d& N+ _/ kHaruhiko Ohashi .................................................................................. 91
: f' [ V# L2 L7 b3 W8 i$ c& R# y11 Direct Cloning of Full-Length Cell Differentially Expressed Genes
, N" \7 {& b; q6 Oby Multiple Rounds of Subtractive Hybridization
" k1 I3 R( u1 @* w! ^Based on Long-Distance PCR and Magnetic Beads
: D+ M3 T! B1 h' g+ BXin Huang, Zhenglong Yuan, and Xuetao Cao ................................ 99 T1 c' Y' x! R: M7 g
PART II. CLONING PCR PRODUCTS2 }$ ^7 z" o3 m! y1 w9 u
12 Cloning PCR Products: An Overview- y" o$ \+ f! g! y" e
Baotai Guo and Yuping Bi ................................................................ 1117 N, O+ P- j4 }( }# [7 v$ o4 j, J0 P% r
13 Using T4 DNA Polymerase to Generate Clonable PCR Products
6 V+ c/ D* h5 y" Q# M1 GKai Wang ............................................................................................. 121
5 a+ u+ l) F0 i# Q% x14 Enzyme-Free Cloning of PCR Products
! E, @3 ~" t( F: h, Zand Fusion Protein Expression
( D6 }4 ^: W" Y# O: {3 c$ ?Brett A. Neilan and Daniel Tillett ..................................................... 125
- B7 v+ V. }' T0 o) ]$ b15 Directional Restriction Site-Free Insertion of PCR Products2 ?5 f- p( _( W% c2 J; j! t
into Vectors% q' z! i/ C1 ?3 E! t2 I0 y* a
Guo Jun Chen .................................................................................... 1332 ^4 m& k) F% v/ S/ ?
16 Autosticky PCR:' R* R. s$ i* t1 H- _& W
Directional Cloning of PCR Products with Preformed 5' Overhangs& ^& } f! S6 P8 y9 c
József Gál and Miklós Kálmán......................................................... 141$ R- H1 C. H# }
17 A Rapid and Simple Procedure for Direct Cloning: A2 s0 _$ j; b( q2 \: c( O+ `0 s
of PCR Products into Baculoviruses
# Q2 F M: W Q$ j/ b: ]Tamara S. Gritsun, Michael V. Mikhailov,5 s2 K) c& x/ M/ P% U1 \' r
and Ernest A. Gould ...................................................................... 153
. K( f2 B8 w* z+ o' ]2 N9 `- _PART III. MUTAGENESIS AND RECOMBINATION. Q& q4 p0 Y# n- ~& a
18 PCR Approaches to DNA Mutagenesis and Recombination:
/ j0 A0 I9 E! @* o8 y u9 E# ^$ wAn Overview- I0 W, J% b2 Z$ ?5 x A( O- h
Binzhang Shen ................................................................................... 167' A/ F7 t( x3 [6 z
19 In-Frame Cloning of Synthetic Genes Using PCR Inserts
: e- S& d, n6 rJames C. Pierce ................................................................................. 1750 J7 q; F0 s0 T5 M6 D! ~
20 Megaprimer PCR
4 l$ ^& [7 i' W6 z! t: t5 MSailen Barik ........................................................................................ 189& W) U" Y. _) l/ M6 W) b5 F
21 PCR-Mediated Recombination:
0 n2 B+ M- A) p- |" c/ CA General Method Applied to Construct Chimeric Infectious$ N8 E( X6 r& a7 {1 e' p
Molecular Clones
3 R! \4 _; s% Q4 j1 S& DGuowei Fang, Barbara Weiser, Aloise Visosky, Timothy Moran,
( S8 n1 |# R. i. J8 fand Harold Burger ......................................................................... 197) G& {' F3 F( e& T' I: f T
22 PCR Method for Generating Multiple Mutations at Adjacent Sites
2 g7 [, y% V$ Y" G: ~Jiri Adamec ......................................................................................... 207
3 c$ e) F" Z7 l L2 ]6 y$ t, V23 A Fast Polymerase Chain Reaction-Mediated Strategy for Introducing
, ^. r$ f- q0 g/ Z) v1 dRepeat Expansions into CAG-Repeat Containing Genes/ S2 t2 x7 U5 u+ R" z1 F. j
Franco Laccone ................................................................................. 217/ b8 a9 q9 x; M! a
24 PCR Screening in Signature-Tagged Mutagenesis of Essential Genes
' F# O# ]- ?* {* ^: X5 h. f3 TDario E. Lehoux and Roger C. Levesque ....................................... 225
1 g: P+ H, G/ ~+ U4 j1 f1 d25 Staggered Extension Process (StEP) In Vitro Recombination
; K. X6 X# O" z4 ^; \7 `5 mAnna Marie Aguinaldo and Frances Arnold ................................... 235
- q ~& t1 t7 a) e2 f6 E26 Random Mutagenesis by Whole-Plasmid PCR Amplification V7 \7 Q- J) `9 D$ b! J+ S
Donghak Kim and F. Peter Guengerich .......................................... 241* E4 _* [6 B; x+ S
PART IV. CLONING UNKNOWN NEIGHBORING DNA- e4 U# |- }4 s* y& J( w
27 PCR-Based Strategies to Clone Unknown DNA Regions
$ @- V6 |- A2 Q+ |* f( efrom Known Foreign Integrants: An Overview
4 x6 K; u& d/ V& |/ d9 T- J1 D& yEric Ka-Wai Hui, Po-Ching Wang, and Szecheng J. Lo ................ 249
! u, r4 }& g+ G9 _) G28 Long Distance Vectorette PCR (LDV PCR)
- P4 S- _: o1 }% S, e" UJames A. L. Fenton, Guy Pratt, and Gareth J. Morgan ................. 275- p# b, Q" s/ [
29 Nonspecific, Nested Suppression PCR Method. N$ t L( P% ]. X5 S, J
for Isolation of Unknown Flanking DNA (“Cold-Start Method”)+ ?: I9 T7 E3 W
Michael Lardelli .................................................................................. 285. t) |* x8 c' x
30 Inverse PCR: cDNA Cloning
. @3 a5 l) A c$ p* g4 TSheng-He Huang ................................................................................ 293" G/ \. J6 A, t% \- {9 u
31 Inverse PCR: Genomic DNA Cloning9 z& S; _" P0 ]. ~: ]( r+ x
Ambrose Y. Jong, Anna T’ang, De-Pei Liu,
8 \; e2 {# {5 oand Sheng-He Huang .................................................................... 301. _1 r! q) ~0 F5 q; G" U1 [
32 Gene Cloning and Expression Profiling by Rapid Amplification; N" D7 E+ S- M F! |; n' d
of Gene Inserts with Universal Vector Primers
9 Z+ ?, i; @. D: l: W8 J+ G/ d: bSheng-He Huang, Hua-Yang Wu, and Ambrose Y. Jong .............. 309* ~( P! J' |( M& E0 k
33 The Isolation of DNA Sequences Flanking Tn5 Transposon Insertions/ [# ?! r1 G) j+ A# j" m, ?
by Inverse PCR/ c9 r5 |; G9 `. s% j* G: e2 E7 S
Vincent J. J. Martin and William W. Mohn ...................................... 315- \4 H1 a; ~: Q [
34 Rapid Amplification of Genomic DNA Sequences Tagged
2 s w8 T6 S' A4 _2 r, p8 Q2 e+ p' hby Insertional Mutagenesis
. B6 ?$ T' ^4 Q6 S1 N) LMartina Celerin and Kristin T. Chun ................................................ 325
. D1 @% X+ }" m/ ?$ p35 Isolation of Large Terminal Sequences of BAC Inserts Based
* K* V- Y. m$ A' Lon Double-Restriction-Enzyme Digestion Followed
) x8 o" e" E8 O* ?+ Bby Anchored PCR7 N2 l8 B9 d% N. S) h; h
Zhong-Nan Yang and T. Erik Mirkov ............................................... 337
- Q9 B; r) ^9 K) X36 A “Step Down” PCR-Based Technique for Walking; z* Y- ^8 s, w
Into and the Subsequent Direct Sequence Analysis3 q: K8 |6 `6 J
of Flanking Genomic DNA- A, {! U1 f: R+ S4 o
Ziguo Zhang and Sarah Jane Gurr .................................................. 343; i5 @4 h3 l4 N6 `% Y, i9 z
PART V. LIBRARY CONSTRUCTION AND SCREENING
5 E5 c8 X4 b- ~37 Use of PCR in Library Screening: An Overview( P! p+ J V% f) E
Jinbao Zhu .......................................................................................... 353
4 C. T& h* r! @38 Cloning of Homologous Genes by Gene-Capture PCR8 r. d h U f0 x3 T
Renato Mastrangeli and Silvia Donini ............................................. 359
# `3 x7 L- O) ?9 I, ^5 d" |. _39 Rapid and Nonradioactive Screening of Recombinant Libraries by PCR9 Y* m9 [$ d$ Q* y2 d. [
Michael W. King ................................................................................. 377
) Q. E: w4 J+ D. N+ X40 Rapid cDNA Cloning by PCR Screening (RC-PCR). ?: ?$ v0 [8 q& V3 l* H7 \
Toru Takumi ....................................................................................... 385( @" } D1 h& H6 J4 K( E+ s
41 Generation and PCR Screening of Bacteriophage λ Sublibraries2 H+ N- w* h5 y1 P5 A' Q+ z
Enriched for Rare Clones (the “Sublibrary Method”)
6 p1 q# B- @3 a- oMichael Lardelli .................................................................................. 391 T) c4 A+ N8 v z9 \0 \7 b
42 PCR-Based Screening for Bacterial Artificial Chromosome Libraries
/ x2 k8 ?& @1 C2 b1 {Yuji Yasukochi ................................................................................... 4019 L; X+ D8 j, n$ ^
43 A 384-Well Microtiter-Plate-Based Template Preparation
+ ~4 E: y) C3 I: u5 \and Sequencing Method# s! Y, _ u( F* f6 k" F3 L X; p
Lei He and Kai Wang ......................................................................... 411
* M# @# \: L9 h! x44 A Microtiter-Plate-Based High Throughput PCR Product3 Z+ e$ D: g1 s: g( u
Purification Method3 |1 S; @! m ^! r
Ryan Smith and Kai Wang ................................................................ 417, y$ d" c V3 Z/ C9 ?
Index ............................................................................................................ 423
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发表于 2010-9-19 18:24
