In vitro cloning of complex mixtures of DNA on microbeads: physical separation of differentially expressed cDNAs.

ABSTRACT: We describe a method for cloning nucleic acid molecules onto the surfaces of 5-micrometer microbeads rather than in biological hosts. A unique tag sequence is attached to each molecule, and the tagged library is amplified. Unique tagging of the molecules is achieved by sampling a small fraction (1%) of a very large repertoire of tag sequences. The resulting library is hybridized to microbeads that each carry approximately 10(6) strands complementary to one of the tags. About 10(5) copies of each molecule are collected on each microbead. Because such clones are segregated on microbeads, they can be operated on simultaneously and then assayed separately. To demonstrate the utility of this approach, we show how to label and extract microbeads bearing clones differentially expressed between tw

SUBMITTER: Brenner S 

PROVIDER: S-EPMC26493 | biostudies-literature | 2000 Feb

REPOSITORIES: biostudies-literature