Large-scale sequencing library production: an adaptor-based strategy.
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).
Povinelli
C M
CM
Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.
Gibbs
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RA
D001483:Base Sequence; D001709:Biotechnology; D015704:CD4 Antigens / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D015723:Gene Library; D005821:Genetic Techniques; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*; D012985:Software Design
D015704:CD4 Antigens; D004247:DNA