UID:
almahu_9949251232502882
Format:
1 online resource (385 p.)
Edition:
1st ed. 1993.
ISBN:
1-59259-510-3
Series Statement:
Methods in Molecular Biology, 23
Content:
The purpose of DNA Sequencing Protocols is to provide detailed practical procedures for the widest range of DNA sequencing meth ods, and we believe that all the vanguard techniques now being applied in this fast-evolving field are comprehensively covered. Sequencing technology has advanced at a phenomenal rate since the original methods were first described in the late 1970s and there is now a huge variety of strategies and methods that can be employed to determine the sequence of any DNA of interest. More recently, a large number of new and innovative sequencing techniques have been developed, including the use of such novel polymerases as Tag poly merase and Sequenase, the harnessing of PCR technology for linear amplification (cycle) sequencing, and the advent of automated DNA sequencers. DNA sequencing is surely one of the most important techniques in the molecular biology laboratory. Sequence analysis is providing an increasingly useful approach to the characterization of biological systems, and major multinational projects are already underway to map and sequence the entire genome of organisms, such as Escherichia coli, Saccharomyces cerevisiae, Caenorhabditis elegans, and Homo sapiens. Most scientists recognize the importance of DNA sequence data and perceive DNA sequencing as a valuable and indispensable aspect of their work. Recent technological advances, especially in the area of automated sequencing, have removed much of the drudg ery that was formerly associated with the technique, whereas innova tive computer software has greatly simplified the analysis and manipulation of sequence data.
Note:
Description based upon print version of record.
,
DNA Sequencing -- M13 Cloning Vehicles -- Cloning into M13 -- Transfection of E. coli with M13 DNA -- M13 Phage Growth and Single-Strand DNA Preparation -- M13 Phage Growth and DNA Purification Using 96 Well Microtiter Trays -- Generation of Random Fragments by Sonication -- Generation of a Nested Set of Deletions Using Exonuclease III -- Sequential Deletions of Single-Stranded DNA -- Subcloning for DNA Sequencing -- Sequencing Using Custom Designed Oligonucleotides -- Dideoxy Sequencing Reactions Using Klenow Fragment DNA Polymerase 1 -- Dideoxy Sequencing Reactions Using T7 Polymerase -- Dideoxy Sequencing Reactions Using Sequenase Version 2.0 -- Dideoxy Sequencing Reactions Using Taq Polymerase -- Pouring Linear and Buffer-Gradient Sequencing Gels -- Electrophoresis of Sequence Reaction Samples -- Plasmid Sequencing -- Plasmid Sequencing -- Direct Sequencing of Inserts Cloned into Lambda Vectors -- Cosmid Sequencing -- Genomic Sequencing -- Sequencing of Double-Stranded PCR Products -- Sequencing Double-Stranded Linear DNA with Sequenase and [?-35S] d.ATP -- Solid Phase PCR Sequencing of Biotinylated Products -- Cycle Sequencing -- Direct Blotting Electrophoresis -- Multipllex DNA Sequencing -- DNA Sequencing by Chemiluminescent Detection -- Reverse Sequencing of M13 Cloned DNA -- Terminal Labeling of DNA for Maxam and Gilbert Sequencing -- DNA Sequencing the Chemical Method -- DNA Sequencing by Chemical Degradation Using One, Two, and Four Different Fluorophores -- Linear Amplification Sequencing with Dye Terminators -- Sequencing Reactions for the Applied Biosystems 373AAutomated DNA Sequencer -- Sequencing Reactions for ALF (EMBL) Automated DNA Sequencer -- Sequencing Using the Du Pont Genesis™ 2000 DNA Analysis System -- The Use of Robotic Workstations in DNA Sequencing.
,
English
Additional Edition:
ISBN 1-4899-3996-2
Additional Edition:
ISBN 0-89603-248-5
Language:
English
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