PCR Sequencing Protocols

Bibliographic Details
Corporate Author: SpringerLink (Online service)
Other Authors: Rapley, Ralph (Editor)
Summary:XI, 221 p. 24 illus.
text
Language:English
Published: Totowa, NJ : Humana Press : Imprint: Humana, 1996.
Edition:1st ed. 1996.
Series:Methods in Molecular Biology, 65
Subjects:
Online Access:https://doi.org/10.1385/0896033449
Format: Electronic Book

MARC

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245 1 0 |a PCR Sequencing Protocols  |h [electronic resource] /  |c edited by Ralph Rapley. 
250 |a 1st ed. 1996. 
264 1 |a Totowa, NJ :  |b Humana Press :  |b Imprint: Humana,  |c 1996. 
300 |a XI, 221 p. 24 illus.  |b online resource. 
336 |a text  |b txt  |2 rdacontent 
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490 1 |a Methods in Molecular Biology,  |x 1940-6029 ;  |v 65 
505 0 |a Preparation and Analysis of DNA Sequencing Gels -- Purification of PCR Products from Agarose Gels for Direct Sequencing -- Enzymatic Fluorescence and Biotin Labeling of Primers for PCR Sequencing -- Direct Sequencing of Double-Stranded PCR Products with the Sequenase Kit and [?-35 S] dATP -- Direct Sequencing by Thermal Asymmetric PCR -- Rapid Sequencing of cDNA Clones Direct Sequencing Using Sequential Linear/Asymmetric PCR -- Direct Sequencing of PCR Products Using Chemiluminescent Detection -- Direct DNA Sequencing of PCR Products Using Magnetic Beads -- Affinity Capture and Solid-Phase Sequencing of Biotinylated PCR Products -- Analysis of Nucleotide Sequence Variations by Solid-Phase Minisequencing -- Nonradioactive PCR Sequencing Using Digoxigenin -- Silver Sequencing™ -- Direct Sequencing of PCR Products with DNA-Binding Proteins -- PCR Sequencing with the Aid of Detergents -- Direct Sequencing with Highly Degenerate and lnosine-Containing Primers -- Determination of Unknown Genomic Sequences Without Cloning -- DNA Sequencing by the Chemical Method -- Direct PCR Sequencing with Denaturants (Formamide) -- Efficient PCR Production of Single-Stranded DNA Sequencing Templates -- Preparation and Direct Automated Cycle Sequencing of PCR Products -- Solid-Phase Automated Sequencing of PCR-Amplified Genomic DNA -- Cloning PCR Products for Sequencing in M13 Vectors -- Sequencing PCR Products Cloned into M13 Vectors -- Genomic Amplification with Transcript Sequencing (GAWTS) -- DNA Rescue by the Vectorette Method -- Sequencing of (dA:dT) Cloned Mixed PCR Products from Microbial Populations. 
520 |a Advances in bioscience research usually arise as a result of the continu­ ing refinement of existing technologies. However, there are a number of occa­ sions v^rhere newly developed methodologies have a profound effect on nearly all areas of research. Frequently these are techniques that are elegantly simple in concept and require minimal technical manipulation. Two of these revolu­ tionary techniques are the focus ofPCR Sequencing Protocols. The first such technique is enzymatic chain termination sequencing developed by Sanger and his co-workers in Cambridge and reported in 1977. This essentially brought the possibility of deriving nucleotide sequence information in a very short time scale and has been widely accepted in many laboratories as a routine molecular biological research tool. Furthermore, it has not only led to the sequencing of many genes and gene fragments, but has also allowed the tech­ nical means of sequencing the human genome. The second technique that has found widespread acceptance in basic applied research and many routine applications is the polymerase chain reac­ tion. This technique, first reported in 1985 by MuUis and his colleagues, pro­ vides the means to amplify nucleic acid sequence, which immediately proved invaluable in nearly all fields of biological laboratory research. Here, as with enzymatic DNA sequencing, is a very simple concept that relies on minimal information to prepare short oligonucleotide primers that direct the synthesis of a specified fi-agment o f DNA in the presence of a thermostable DNA polymerase. 
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