PCR Sequencing Protocols

Häftad, Engelska, 2013

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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.

Produktinformation

Utgivningsdatum2013-08-13
Mått152 x 229 x 14 mm
Vikt354 g
FormatHäftad
SpråkEngelska
SerieMethods in Molecular Biology
Antal sidor221
FörlagHumana Press Inc.
ISBN9781489940384

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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.

...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