E-Book Overview
DNA Sequencing Protocols provides you with the knowledge to become a sequencing expert. An all-star cast of investigators covers virtually all aspects, including developments in cycle sequencing, sequencing PCR products, sequencing lambda and cosmids, multiplex sequencing, direct blotting electrophoresis, sequencing of chemiluminescence, and automated sequencing.
E-Book Content
CHAIVER
1
DNA Sequencing Hugh G. Grifin
and Annette M. Grifin
1. Introduction Methods to determine the sequenceof DNA were developed in the late 1970s (1,2) and have revolutionized the science of molecular genetics. The DNA sequences of many different genes from diverse sources have been determined, and the information is stored in international databanks such as EMBL, GenBank, and DDBJ. Many scientists now accept that sequence analysis will provide an increasingly useful approach to the characterization of biological systems. Projects are already underway to map and sequencethe entire genome of organisms such as Escherichia coli, Saccharomyces cerevisiae, Caenorhabditis elegans, and Homo sapiens. In the recent past, large-scale sequencing projects such as these were often dismissed as prohibitively expensive and of little short-term benefit, while DNA sequencing itself was seen as a repetitive and unintellectual pursuit. However, this view is now changing and most scientists recognize the importance of DNA sequence data and perceive DNA sequencing as a valuable and often indispensable aspect of their work. Recent technological advances, especially in the area of automated sequencing, have removed much of the drudgery that used to be associated with the technique, and modern innovative computer software has greatly simplified the analysis and manipulation of sequence data. Large-scale sequencing From Methods m Molecular Biology, Vol. 23. DNA Sequencmg Protocols Edi