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

C M Povinelli; R A Gibbs

doi:10.1006/abio.1993.1144

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

C M Povinelli·R A Gibbs

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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).

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Base 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 Journal Article Research Support, Non-U.S. Gov't

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C M Povinelli, R A Gibbs. 1993. Large-scale sequencing library production: an adaptor-based strategy.. https://doi.org/10.1006/abio.1993.1144

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