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Contents
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About this book
A superb, comprehensive collection of the most reliable protocols for PCR sequencing - all developed and described by eminent researchers and scientists who use them successfully every day. These protocols deal practically with those problems accompanying the rapid reassociation of amplified complementary strands by showing how sequencing reactions may be effectively modified, as well as complex methods of primer manipulation that enable the affinity purification, or its transcript production, of homogeneous single strands. They also provide all the necessary details of PCR purification, primer labeling, the preparation of sequencing gels, as well as of conventional methods for cloning PCR products into vectors and their subsequent sequencing.
Contents
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 Sequencase Kit and [a-35S] 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 Biotinylated PCR Products. Analysis of Nucleotide Sequence Variations by Solid-Phase Minisequencing. Nonradioactive PCR Sequencing Using Digoxygenin. Silver SequencingTM: Nonradioactive Cycle Sequencing of dsDNA. Direct Sequencing of PCR Products with DNA-Binding Proteins. PCR Sequencing with the Aid of Detergents. Direct Sequencing with Highly Degenerate and Inosine- 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. Genome Amplification with Transcript Sequencing (GAWTS). DNA Rescue by Vectorette Method. Sequencing (dAidT) of Cloned Mixed PCR Products from Microbial Populations. Index.
Customer Reviews
Handbook / Manual
Out of Print
Edited By: Ralph Rapley
221 pages, Figs, tabs
...this is a good collection of well presented PCR protocols that should find its way into many laboratories...if you are planning on performing a large amount of PCR sequencing, or have been having problems sequencing PCR products, this book has a procedure or advice that will help. - FEBS Letters The book is a must for investigators in molecular biology and involved with sequencing and setting up PCR protocols. I highly recommend this most comprehensive volume for all molecular biologists. - SMG Quarterly