Abstract
The PCR method has provided a substitute for most of the repetitive types of molecular cloning and template preparation for sequencing. In combination with automated sequencing techniques, PCR will provide the fastest and most efficient means of generating nucleotide acid sequence information. The purpose of this chapter is to review methods for preparation of sequencing templates and performing direct sequencing of PCR products.
Keywords
Sequencing by ligationDNA sequencingTemplateComputational biologyCloning (programming)DNA nanoball sequencingBiologyMassive parallel sequencingDNAGeneticsComputer scienceGenomic libraryBase sequence
Related Publications
Ligation-independent cloning of PCR products (LIC-PCR)
A new procedure has been developed for the efficient cloning of complex PCR mixtures, resulting in libraries exclusively consisting of recombinant clones. Recombinants are gener...
Cell-free cloning using φ29 DNA polymerase
We describe conditions for rolling-circle amplification (RCA) of individual DNA molecules 5–7 kb in size by >10 9 -fold, using φ29 DNA polymerase. The principal difficulty wi...
Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis.
Fluorescent dye-labeled DNA primers have been developed that exploit fluorescence energy transfer (ET) to optimize the absorption and emission properties of the label. These pri...
Accurate Multiplex Polony Sequencing of an Evolved Bacterial Genome
We describe a DNA sequencing technology in which a commonly available, inexpensive epifluorescence microscope is converted to rapid nonelectrophoretic DNA sequencing automation....
Direct Cloning and Sequence Analysis of Enzymatically Amplified Genomic Sequences
A method is described for directly cloning enzymatically amplified segments of genomic DNA into an M13 vector for sequence analysis. A 110-base pair fragment of the human β-glob...
Publication Info
- Year
- 1989
- Type
- book-chapter
- Pages
- 45-60
- Citations
- 96
- Access
- Closed
External Links
Social Impact
Altmetric
PlumX Metrics
Social media, news, blog, policy document mentions
Citation Metrics
96
OpenAlex
Cite This
Ulf Gyllensten
(1989).
Direct Sequencing of In Vitro Amplified DNA.
Palgrave Macmillan UK eBooks
, 45-60.
https://doi.org/10.1007/978-1-349-20235-5_5
Identifiers
- DOI
- 10.1007/978-1-349-20235-5_5