Large-scale sequencing library production: an adaptor-based strategy.

TitleLarge-scale sequencing library production: an adaptor-based strategy.
Publication TypeJournal Article
Year of Publication1993
AuthorsPovinelli, CM, Gibbs, RA
JournalAnal Biochem
Volume210
Issue1
Pagination16-26
Date Published1993 Apr
ISSN0003-2697
KeywordsBase Sequence, Biotechnology, CD4 Antigens, Cloning, Molecular, DNA, Evaluation Studies as Topic, Gene Library, Genetic Techniques, Genetic Vectors, Humans, Molecular Sequence Data, Sequence Analysis, DNA, Software Design
Abstract

A significant bottleneck in large-scale DNA sequencing projects is the creation of high-quality random libraries suitable for sequencing. We have utilized an adaptor-based strategy and developed a simple protocol to reliably generate single-insert random subclone M13 libraries yielding uniform coverage and > 10(3) clones per microgram of DNA. Of the resulting white plaques, 95% contained inserts, < 0.5% were due to adaptors with no insert, and none of the clones were found to contain extraneous DNA (0/600) or multiple inserts (0/300).

Alternate JournalAnal. Biochem.
PubMed ID8489013