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090727 ||| eng |
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|a 9781592595532
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| 050 |
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4 |
|a QD415-436
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| 100 |
1 |
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|a White, Bruce A.
|e [editor]
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| 245 |
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|a PCR Cloning Protocols
|h Elektronische Ressource
|b From Molecular Cloning to Genetic Engineering
|c edited by Bruce A. White
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| 250 |
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|a 1st ed. 1997
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| 260 |
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|a Totowa, NJ
|b Humana Press
|c 1997, 1997
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| 300 |
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|a XIV, 490 p
|b online resource
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| 505 |
0 |
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|a Recombination and Site-Directed Mutagenesis Using Recombination PCR -- In Vitro Recombination and Mutagenesis of DNA -- In-Frame Cloning of Synthetic Genes Using PCR Inserts -- Creation of Chimeric Junctions, Deletions, and Insertions by PCR -- Mutagenesis and Gene Fusion by Megaprimer PCR -- Rapid and Efficient One-Tube PCR-Based Mutagenesis Method -- Thermostable Ligase-Mediated Incorporation of Mutagenic Oligonucleotides During PCR Amplification -- Linker Scanning Mutagenesis by Three-Step PCR -- Sequence Inversion by Flip-PCR -- PCR Site-Directed Mutagenesis Using Pyrococcus sp GB-D Polymerase Coupled to a Rapid Screening Procedure -- Using the SELEX Combinatorial Chemistry Process to Find High Affinity Nucleic Acid Ligands to Target Molecules -- Cloning Unknown Neighboring DNA -- Rapid Amplification of cDNA Ends -- Amplification of Gene-Regulating Regions with Single-Sided Specificity --
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| 505 |
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|a Performing and Optimizing PCR -- PCR -- XL PCR Amplification of Long Targets From Genomic DNA -- Amplification of DNA Sequences Up To 5 kb from Small Amounts of Genomic DNA Using Tub DNA Polymerase -- One-Step Optimization Using Touchdown and Stepdown PCR -- GC-Rich Template Amplification by Inverse PCR -- Coupled One-Step Reverse Transcription and Polymerase Chain Reaction Procedure for Cloning Large cDNA Fragments -- Cloning PCR Products -- Using T4 DNA Polymerase to Generate Clonable PCR Products -- Rapid (Ligase-Free) Subcloning of PCR Products -- Cloning PCR Products Utilizing the T/A Overhang and a Kit -- Cloning Unmodified PCR Products Using Engineered Xcml Restriction Sites in a Portable Cassette -- A T-Linker Strategy for Modification and Directional Cloning of PCR Products -- Recovery of DNA Amplification Products from Silver-Stained Polyacrylamide Gels -- Mutagenesis, Recombination, and In Vitro Selection --
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|a The Use of PCR for Differential Screening of cDNA Libraries -- Identification and Cloning of Differentially Expressed Genes by DDRT-PCR -- Cloning Members of Gene Families -- Cloning Gene Family Members Using PCR with Degenerate Oligonucleotide Primers -- Amplification Using Degenerate Primers with Multiple Inosines to Isolate Genes with Minimal Sequence Similarity -- Designing PCR Primers to Amplify Specific Members or Subgroups of Multigene Families -- Screening Gene Family-Enriched cDNA Sublibraries with an Unamplified cDNA Probe
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| 505 |
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|a An End-Trimming Method and Its Application to Amplify Adjacent cDNA and Genomic DNA Fragments by PCR -- Anchoring a Defined Sequence to the 5? Ends of mRNAs -- Rapid Directional Walk Within DNA Clones by Step-Out PCR -- Inverse PCR -- Rapid Amplification of Gene Ends (RAGE) from Gene Libraries by Anchored PCR -- Isolation of Coding Sequences from Yeast Artificial Chromosome (Yac) -- Library Construction and Screening -- cDNA Libraries from a Low Amount of Cells -- Rapid and Nonradioactive Screening of Recombinant Libraries by PCR -- Use of PCR for cDNA Library Screening -- Generation and PCR Screening of Bacteriophage ? Sublibraries Enriched for Rare Clones (the “Sublibrary Method”) -- Differential and Subtractive Approach By cDNA Analysis and Cloning -- Normalization of cDNA Sequence Representation by Molecular Selection -- Subtractive cDNA Cloning Using Magnetic Beads and PCR -- Generation of a PCR-Renewable Source of Subtractive cDNA --
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| 653 |
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|a Biochemistry
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| 041 |
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7 |
|a eng
|2 ISO 639-2
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| 989 |
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|b SPRPROT
|a Springer Protocols Archive 1981-2004
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| 490 |
0 |
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|a Methods in Molecular Biology
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| 024 |
8 |
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|a 10.1385/0896034836
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| 856 |
4 |
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|u https://doi.org/10.1385/0896034836?nosfx=y
|x Verlag
|3 Volltext
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| 082 |
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|a 572
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| 520 |
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|a The advent of PCR, with its power to amplify tiny amounts of DNA, quickly spawned the development of many analytical procedures that are widely used for detection, measurement, and characterization. However, creative investigators soon discovered the power of PCR for synthetic or preparative uses. This volume focuses on such preparative PCR protocols, which can be used in the cloning and modification of DNA. PCR Cloning Protocols: From Molecular Cloning to Genetic Engineer ing is divided into seven parts, each containing a collection of chapters address ing a general approach or goal. Part I presents basic PCR protocols, emphasizing optimizing conditions for (he amplification of DNA fragments of several kilobases in length. Part 11 offers several procedures for cloning PCR prod ucts, depending on whether a specific restriction site can be used in the clon ing vector, the PCR product is to be gel purified before cloning, or the fragmeni needs to be inserted in one or both orientations. Part III includes several pro tocols involved in the mutagenesis of DNA, either site-directed or not, as well as several approaches to recombinant PCR, either for mutagenesis or building a custom gene, as well as one chapter describing a specific use of in vitro selection. Part IV addresses the frequent need to amplify and clone segments of DNA that are to the right, left, or scattered within a stretch of DNA (either vector, chromosomal, or cDNA) of known sequence
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