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本帖最后由 细胞海洋 于 2010-9-19 20:18 编辑 : @2 ~; x! P; I {, L ]( f
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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，
# }! v5 r7 k0 q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。5 {' E. _7 s: u' F( E' k9 t( J
Preface .............................................................................................................v
H" U" E7 T! {1 n4 `5 dContributors .....................................................................................................xi7 h* h! G" B% h7 W& G- H+ G7 f
PART I. PERFORMING AND OPTIMIZING PCR5 M5 Q* f6 D( }3 `/ N# I) s/ H, Q7 b
1 Polymerase Chain Reaction: Basic Principles and Routine Practice
* {0 N+ Y) @) P% o* nLori A. Kolmodin and David E. Birch .................................................. 3
4 z3 z$ `6 y! ]; r4 a7 j" _( c2 Computer Programs for PCR Primer Design and Analysis: U# D2 L* V/ W6 D# a/ l# o! `' R
Bing-Yuan Chen, Harry W. Janes, and Steve Chen ........................ 199 F+ S. N5 t% V: Y! R; p% G
3 Single-Step PCR Optimization
0 I( W4 J* v4 N5 pUsing Touchdown and Stepdown PCR Programming" L7 T$ c, g0 [+ ?! Y
Kenneth H. Roux .................................................................................. 311 n0 w5 M M! V
4 XL PCR Amplification of Long Targets from Genomic DNA" h- ^& V; |+ ?1 n$ U4 I" h6 G9 }3 i
Lori A. Kolmodin .................................................................................. 372 y5 t7 W' {! S* z
5 Coupled One-Step Reverse Transcription and Polymerase Chain( P. m3 f$ e+ G6 G
Reaction Procedure for Cloning Large cDNA Fragments
9 r9 e, {" K6 |Jyrki T. Aatsinki ................................................................................... 53
- ~$ a+ o. F1 d. d' k6 Long Distance Reverse-Transcription PCR
; T) R; C+ [1 a6 HVolker Thiel, Jens Herold, and Stuart G. Siddell ............................. 59
1 x, C8 i$ I6 [. Q- F3 ~7 Increasing PCR Sensitivity for Amplification
2 I l' X( E* w" [' g/ vfrom Paraffin-Embedded Tissues) ]2 E2 J- x7 A& [# J
Abebe Akalu and Juergen K. V. Reichardt ....................................... 67
* f8 ^+ ~3 L1 z& P, N" d8 `6 x8 GC-Rich Template Amplification by Inverse PCR:
z' P. X. Q* E8 R/ |DNA Polymerase and Solvent Effects* Y6 W' m# \1 P1 b1 {
Alain Moreau, Da Shen Wang, Steve Forget, Colette Duez,6 m: {+ m# n6 M& ~ p6 J3 k: \
and Jean Dusart............................................................................... 75" m4 X0 _6 |7 p/ Q d; O5 X
9 PCR Procedure for the Isolation of Trinucleotide Repeats+ @4 ?7 b& P! f: A
Teruaki Tozaki ...................................................................................... 81
& D0 {9 F/ v# D/ q$ r4 X10 Methylation-Specific PCR% @5 l, `" x& s- C" w* c
Haruhiko Ohashi .................................................................................. 91
: m) k: `6 B L9 z9 B z11 Direct Cloning of Full-Length Cell Differentially Expressed Genes
. q7 R8 s0 c' V) kby Multiple Rounds of Subtractive Hybridization& m: ?% p4 U0 V+ c, \' |, b8 m6 V2 L
Based on Long-Distance PCR and Magnetic Beads4 S) P1 g7 k9 ~; U$ n7 a# f |
Xin Huang, Zhenglong Yuan, and Xuetao Cao ................................ 99
* h3 n* `% K' APART II. CLONING PCR PRODUCTS
: }/ g+ u, V% B! q% f12 Cloning PCR Products: An Overview
K( i7 Q7 c H- O5 HBaotai Guo and Yuping Bi ................................................................ 111
; H# [+ R( p# q) ]/ m. P; C0 V13 Using T4 DNA Polymerase to Generate Clonable PCR Products/ i$ v# F( A9 ^0 x
Kai Wang ............................................................................................. 121
3 G2 K7 `1 s$ N0 b+ n14 Enzyme-Free Cloning of PCR Products& m( w4 o {$ `( R
and Fusion Protein Expression# f/ h4 F# i* Z V
Brett A. Neilan and Daniel Tillett ..................................................... 125/ X! k( E1 g! |" B R
15 Directional Restriction Site-Free Insertion of PCR Products/ ?# c6 S% f- X# ?9 b
into Vectors: K& G% m- u K5 s% o# ]
Guo Jun Chen .................................................................................... 133/ q$ o4 m- N0 O6 B F. i7 U
16 Autosticky PCR:" H v! q5 Z, R4 c% c$ s
Directional Cloning of PCR Products with Preformed 5' Overhangs
/ L. {# _$ t1 m3 cJózsef Gál and Miklós Kálmán......................................................... 141
# f: W6 J# l. `: a1 q' z* [% o17 A Rapid and Simple Procedure for Direct Cloning
/ S5 E4 D- y* K oof PCR Products into Baculoviruses! ]; ~7 T }( m" h8 J n" h ]
Tamara S. Gritsun, Michael V. Mikhailov,
& d5 y& c8 ]$ z# Q* }6 ?) O% Tand Ernest A. Gould ...................................................................... 1536 |9 b+ _3 b! R! z2 n2 q
PART III. MUTAGENESIS AND RECOMBINATION# q% x1 Z" v4 ?
18 PCR Approaches to DNA Mutagenesis and Recombination:
# o6 I' p% {3 ^# pAn Overview4 D! y- i$ ~ l7 P
Binzhang Shen ................................................................................... 1672 ?: W& C4 O2 K, N
19 In-Frame Cloning of Synthetic Genes Using PCR Inserts. z3 c" U& E9 U6 n. |0 B) }
James C. Pierce ................................................................................. 175 N% i4 X6 d: F9 |9 d
20 Megaprimer PCR8 u' |- l; b' E4 ~, ~
Sailen Barik ........................................................................................ 189
, d+ x0 a$ T& H21 PCR-Mediated Recombination:
- v5 Y2 Y* K: z I1 y' XA General Method Applied to Construct Chimeric Infectious: {& @: w( U2 I, E0 e
Molecular Clones
5 [; L1 h/ `. U" I" }% F5 hGuowei Fang, Barbara Weiser, Aloise Visosky, Timothy Moran,
/ j: E$ h0 p6 A- J- L6 W+ Gand Harold Burger ......................................................................... 1973 Z4 b' w+ u; W# i
22 PCR Method for Generating Multiple Mutations at Adjacent Sites; b1 E) f' D8 {/ w
Jiri Adamec ......................................................................................... 2072 v; m$ R8 r% W7 ^4 R
23 A Fast Polymerase Chain Reaction-Mediated Strategy for Introducing
% s; V, ~% U8 K2 }8 L1 V* ?Repeat Expansions into CAG-Repeat Containing Genes& x# H3 k+ c* a
Franco Laccone ................................................................................. 217# \: @% w) O+ Q: Q
24 PCR Screening in Signature-Tagged Mutagenesis of Essential Genes
# l ]: }& q+ x8 T8 i x1 z0 CDario E. Lehoux and Roger C. Levesque ....................................... 225 `( Q9 K& ?0 i6 T4 a
25 Staggered Extension Process (StEP) In Vitro Recombination1 x. A' C1 b- @- `, x' [- A2 g1 h! x
Anna Marie Aguinaldo and Frances Arnold ................................... 235) w3 Y/ c; D3 Z/ n$ j1 n3 D* x
26 Random Mutagenesis by Whole-Plasmid PCR Amplification7 C8 ]% }2 a- I3 l$ Q4 n
Donghak Kim and F. Peter Guengerich .......................................... 241* i" S1 k7 X% h1 }# T
PART IV. CLONING UNKNOWN NEIGHBORING DNA! l* C i2 P: D& d, n' ~$ U
27 PCR-Based Strategies to Clone Unknown DNA Regions* e2 |9 T3 Y5 j a
from Known Foreign Integrants: An Overview
: [$ j; |) G$ s$ |/ v* P# }Eric Ka-Wai Hui, Po-Ching Wang, and Szecheng J. Lo ................ 2492 A" t- A _% o
28 Long Distance Vectorette PCR (LDV PCR)
# E% ^1 m8 D3 \7 o5 M a9 @9 l' Q0 wJames A. L. Fenton, Guy Pratt, and Gareth J. Morgan ................. 275" E- p4 S8 y$ k# [, |- u8 |
29 Nonspecific, Nested Suppression PCR Method
- b: P7 W! O% q& @! Y. vfor Isolation of Unknown Flanking DNA (“Cold-Start Method”)
, k1 T; I" ?, u0 z3 r+ m' mMichael Lardelli .................................................................................. 2856 P% z- |& _; l8 h( D& F
30 Inverse PCR: cDNA Cloning' W" d; U0 ?; a+ Y: F+ F2 \
Sheng-He Huang ................................................................................ 293: J2 R8 ^0 w) {! L+ [5 C$ H7 c
31 Inverse PCR: Genomic DNA Cloning$ u; w7 R2 U: ^( t( @) A% l' P
Ambrose Y. Jong, Anna T’ang, De-Pei Liu,/ x% x. @# R. T; ^* N0 {
and Sheng-He Huang .................................................................... 301
. k. y- f( G) r: x2 p7 O32 Gene Cloning and Expression Profiling by Rapid Amplification
+ l( ^1 y% V: i7 O4 t8 yof Gene Inserts with Universal Vector Primers4 e6 r( c2 s$ S
Sheng-He Huang, Hua-Yang Wu, and Ambrose Y. Jong .............. 309
! n( w: K; n( o$ N* @- l, L33 The Isolation of DNA Sequences Flanking Tn5 Transposon Insertions& J: U5 P/ k3 u8 m. n
by Inverse PCR
% h- b3 j1 S( m1 K6 NVincent J. J. Martin and William W. Mohn ...................................... 315
/ c" ?0 m: _1 I0 s0 E34 Rapid Amplification of Genomic DNA Sequences Tagged- p0 M) M r4 {
by Insertional Mutagenesis& A& J$ A& T# ^' z& n7 ~9 _
Martina Celerin and Kristin T. Chun ................................................ 3257 o, w( ^8 q% }
35 Isolation of Large Terminal Sequences of BAC Inserts Based
' p; u- u ]8 u: r& r& K. n# uon Double-Restriction-Enzyme Digestion Followed
* N, R+ l6 q7 {: u# ?! aby Anchored PCR
, V( ~1 T+ y* u6 w6 iZhong-Nan Yang and T. Erik Mirkov ............................................... 337
) K3 J J2 t: y) k36 A “Step Down” PCR-Based Technique for Walking
- t7 t! w6 ^3 R5 @ {/ mInto and the Subsequent Direct Sequence Analysis' C. I1 W1 e- q2 h4 d
of Flanking Genomic DNA
0 h$ u" v% K* F! j# jZiguo Zhang and Sarah Jane Gurr .................................................. 343
# T" F7 T9 T. F7 m7 r: SPART V. LIBRARY CONSTRUCTION AND SCREENING
; I6 `: R9 C8 f$ W; ~% _37 Use of PCR in Library Screening: An Overview; j& y/ K/ ^7 j- V) Y9 ?
Jinbao Zhu .......................................................................................... 353
6 d" |8 v! z3 k# @& H9 B0 ], d38 Cloning of Homologous Genes by Gene-Capture PCR
! n: r3 Q4 a; j3 w/ b% {/ t' M) u6 ?! FRenato Mastrangeli and Silvia Donini ............................................. 359
+ |) \* z- {" c) l# L5 C5 F, L- J% S39 Rapid and Nonradioactive Screening of Recombinant Libraries by PCR
5 ?: N' o3 r: w: cMichael W. King ................................................................................. 377
. {3 h$ D7 K/ G( |# z) S! t x40 Rapid cDNA Cloning by PCR Screening (RC-PCR)
6 b, Y, s" p0 s2 N6 J' ]: W3 qToru Takumi ....................................................................................... 385" t& |& f, U: d% w
41 Generation and PCR Screening of Bacteriophage λ Sublibraries
1 {; l( s# v4 G8 REnriched for Rare Clones (the “Sublibrary Method”)
% C+ ^6 [( B- R( CMichael Lardelli .................................................................................. 3912 G$ y0 J# }# W1 @! q7 q' o, q
42 PCR-Based Screening for Bacterial Artificial Chromosome Libraries
- u% J, h2 v3 N: `1 fYuji Yasukochi ................................................................................... 4012 [* t1 G: J$ H+ k- B x Z' ?! b
43 A 384-Well Microtiter-Plate-Based Template Preparation: B9 [. J! _5 _6 k
and Sequencing Method
2 ~/ N( X* }, L: nLei He and Kai Wang ......................................................................... 411
& X4 v. y/ b5 k6 H6 S44 A Microtiter-Plate-Based High Throughput PCR Product
6 l( a2 N, ~! T/ D$ y; XPurification Method
2 d+ u5 I6 ^" V) B k* _% ORyan Smith and Kai Wang ................................................................ 417! ~+ z! Z+ k' S5 W& l
Index ............................................................................................................ 423: @8 e: Z' Y4 j' A
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发表于 2010-9-19 18:24
