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
Date Published1993 Apr
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

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

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