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N8029 Sigma

Nucleic Acid Protocols Handbook

  •  ISBN-10 0-89603-841-6

  •  ISBN-13 978-0-89603-841-7

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publication info   R. Rapley, ed., Humana Press, 2000, 1072 pp., soft cover
mfr. no.   Humana

Description

General description

A comprehensive volume of molecular biology methods ranging from DNA extraction to gene localization in situ. The 120 techniques cited list all necessary materials and reagents, step-by-step instruction, pitfalls to avoid, troubleshooting tips, alternate methods, and reasons for certain steps. All key elements contributing significantly to success or failure in the lab. It is a collection of all classic and cutting-edge techniques for isolation, analysis, and manipulation of nucleic acids.

Table of Contents

PART I. Nucleic Acid Extraction.
Isolation of High-Molecular-Weight DNA from Animal Cells, Isolation of mRNA by Affinity Chromatography, Isolation and Purification of DNA from Plants, Purification of Uncontaminated, Intact Plant RNA, Isolating Chromosomal DNA from Bacteria, Bacterial DNA Extraction for Polymerase Chain Reaction and Pulsed-Field Gel Electrophoresis, Isolation of Fungal Nucleic Acids.......................
PART II. Basic Separation and Analysis of DNA.
Restriction Endonuclease Digestion of DNA, Agarose Gel Electrophoresis of Nucleic Acids, Preparation of RNA Dot Blots, Native Polyacrylamide Gel Electrophoresis, Southern Blotting of Agarose Gels by Capillary Transfer, . Pulsed-Field Gel Electrophoresis, HPLC of DNA and PCR Products
PART III. Probe Design, Synthesis, and Labeling.
End-Labeling of DNA Fragments, Nick Translation and Random Hexamer Labeling of DNA, Generation of Labeled Probes by Polymerase Chain Reaction, Preparation of Direct, Enzyme-Labeled DNA Probes, Random Prime Labeling of DNA Probes with Fluorescein-Tagged Nucleotides, Hybridization and Detection of Fluorescein-Labeled DNA Probes Using Chemiluminescence, ..................
PART IV. RNA Analysis Techniques.
Formaldehyde Gel Electrophoresis of Total RNA, . RNA Probes for the Analysis of Gene Expression, Primer Extension Analysis of mRNA, S1 Mapping Using Single-Stranded DNA Probes, Measurements of Rate of Transcription in Isolated Nuclei by Nuclear ¿Run-Off¿ Assay, One-Tube RT-PCR with Sequence-Specific RT Primers, . Characterization of RNA Using Continuous RT-PCR Coupled with ELOSA, ................
PART V. Gene Library Construction and Screening.
Production of Double-Stranded cDNA for Gene Library Synthesis, Using Rapid Amplification of cDNA Ends (RACE) to Obtain Full-Length cDNAs, cDNA Library Construction Using Streptavidin-Paramagnetic Beads and PCR, Rapid (Ligase-Free) Subcloning of Polymerase Chain Reaction Products, Subtraction Hybridization cDNA Libraries, Cloning Polymerase Chain Reaction Products Utilizing the T/A Overhang and a Kit, .................
PART VI. DNA Sequencing.
Preparation and Analysis of DNA Sequencing Gels, DNA Sequencing of Plasmids, Sequencing DNA Fragments Cloned into M13 and Phagemid Vectors, Direct cDNA Sequencing Using Sequential Linear/Asymmetric Polymerase Chain Reaction, . Purification and Enzymatic Sequencing of Polymerase Chain Reaction Products, Direct Polymerase Chain Reaction Sequencing with Denaturants, .....................
PART VII. Basic Polymerase Chain Reaction Methods.
Polymerase Chain Reaction: Basic Principles and Routine Practice, Primer Selection and Design for Polymerase Chain Reaction, One-Step Optimization Using Touchdown and Stepdown Polymerase Chain Reaction, . Cloning Gene Family Members Using Polymerase Chain Reaction with Degenerate Oligonucleotide Primers
PART VIII. Analyzing Genes, Mutations, and Protein Interactions.
Nonradioactive Differential Display of Messenger RNA, Gene Isolation by Exon Trapping, DNA Rescue by the Vectorette Method, Random Amplified Polymorphic DNA (RAPDs), Restriction Fragment Length Polymorphism, Mutation Screening Using PCR-SSCP: Silver Staining and Isotopic Protocols, Analysis of Nucleotide Sequence Variation by Solid-Phase Minisequencing, ........................
PART IX. Mutagenesis, Transcription and Translation In Vitro.
Generating Nested Deletions with Exonuclease III, Primer-Directed Site-Specific Mutagenesis, Site-Directed Mutagenesis Using a Uracil-Containing Phagemid Template, Site-Directed Mutagenesis Using Double-Stranded Plasmid DNA Templates
PART X. Gene Localization, Mapping In Situ, and Bioinformatics.

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