|Title||Large-scale sequencing library production: an adaptor-based strategy.|
|Publication Type||Journal Article|
|Year of Publication||1993|
|Authors||Povinelli, CM, Gibbs, RA|
|Date Published||1993 Apr|
|Keywords||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|
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 Journal||Anal. Biochem.|