PCR cloning protocols / edited by Bing-Yuan Chen and Harry W. Janes.

Includes bibliographical references and index.

Print version record.

Polymerase chain reaction. Basic principles and routine practice / L.A. Kolmodin and D.E. Birch -- Computer programs for PCR primer design and analysis / B.Y. Chen, H.W. Janes and S. Chen -- Single-step PCR optimization using touchdown and stepdown PCR programming / K.H. Roux -- Xl PCR amplification of long targets from genomic DNA / L.A. Kolmodin -- Coupled one-step reverse transcription and polymerase chain reaction procedure for cloning large cDNA fragments / J.T. Aatsinki -- Long distance reverse-transcription PCR / V. Thiel, J. Herold and S.G. Siddell -- Increasing PCR sensitivity for amplification from paraffin-embedded tissues / A. Akalu and J.K. Reichardt -- GC-rich template amplification by inverse PCR. DNA polymerase and solvent effects / A. Moreau [and others] -- PCR procedure for the isolation of trinucleotide repeats / T. Tozaki -- Methylation-specific PCR / H. Ohashi -- Direct cloning of full-length cell differentially expressed genes by multiple rounds of subtractive hybridization based on long-distance PCR and magnetic beads / X. Huang, Z. Yuan and X. Cao -- Cloning PCR products. An overview / B. Guo and Y. Bi -- Using t4 DNA polymerase to generate clonable PCR products / K. Wang -- Enzyme-free cloning of PCR products and fusion protein expression / B.A. Neilan and D. Tillett -- Directional restriction site-free insertion of PCR products into vectors / G.J. Chen -- Autosticky PCR. Directional cloning of PCR products with performed 5' overhangs / J. Gal and M. Kalman -- A rapid and simple procedure for direct cloning of PCR products into baculoviruses / T.S. Gritsun, M.V. Mikhailov and E.A. Gould -- PCR approaches to DNA mutagenesis and recombination. An overview / B. Shen -- In-frame cloning of synthetic genes using PCR inserts / J.C. Pierce -- Megaprimer PCR / S. Barik -- PCR method for generating multiple mutations at adjacent sites / J. Adamec -- A fast polymerase chain reaction-mediated strategy for introducing repeat expansions into cag-repeat containing genes / F. Laccone -- PCR screening in signature-tagged mutagenesis of essential genes / D.E. Lehoux and R.C. Levesque -- Staggered extension process (step) in vitro recombination / A.M. Aguinaldo and F. Arnold -- Random mutagenesis by whole-plasmid PCR amplification / D. Kim and F.P. Guengerich -- PCR-based strategies to clone unknown DNA regions from known foreign integrants. An overview / E.K. Hui, P.C. Wang and S.J. Lo -- Long distance vectorette PCR (ldv PCR) / J.A. Fenton, G. Pratt and G.J. Morgan -- Nonspecific, nested suppression PCR method for isolation of unknown flanking DNA ("cold-start method") / M. Lardelli -- Inverse PCR. cDNA cloning / S.H. Huang -- Inverse PCR. Genomic DNA cloning / A.Y. Jong [and others] -- Gene cloning and expression profiling by rapid amplification of gene inserts with universal vector primers / S.H. Huang, H.Y. Wu and A.Y. Jong -- The isolation of DNA sequences flanking TN5 transposon insertions by inverse PCR / V.J. Martin and W.W. Mohn -- Rapid amplification of genomic DNA sequences tagged by insertional mutagenesis / M. Celerin and K.T. Chun -- Isolation of large-terminal sequences of bac inserts based on double-restriction-enzyme digestion followed by anchored PCR / Z.N. Yang and T.E. Mirkov -- A "step down" PCR-based technique for walking into and the subsequent direct-sequence analysis of flanking genomic DNA / Z. Zhang and S.J. Gurr -- Use of PCR in library screening. An overview / J. Zhu -- Cloning of homologous genes by gene-capture PCR / R. Mastrangeli and S. Donini -- Rapid and nonradioactive screening of recombinant libraries by PCR / M.W. King -- Rapid cDNA cloning by PCR screening (RC-PCR) / T. Takumi -- Generation and PCR screening of bacteriophage lambda sublibraries enriched for rare clones (the "sublibrary method") / M. Lardelli -- PCR-based screening for bacterial artificial chromosome libraries / Y. Yasukochi -- A 384-well microtiter-plate-based template preparation and sequencing method / L. He and K. Wang -- A microtiter-plate-based high throughput PCR product purification method / R. Smith and K. Wang.

Annotation PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.

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