title: A novel system for large-scale sequencing of cDNA by PCR amplification.
abstract: We have developed a method for constructing a library containing the 3' end fragment of cDNA for large-scale sequencing of cDNA clones. The average size of the insert was 270 bp. Cell lysates that carry plasmids having the cDNA insert were subjected to PCR amplification of the cDNA moiety and the products were subjected to sequencing analysis using an autosequencer. With this protocol, sample preparation became a non-limiting step, that allowed us to sequence as many samples as the autosequencer could handle.
authors.affiliation: Institute for Molecular and Cellular Biology, Osaka University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Okubo
authors.forename: N
authors.initials: N
authors.lastname: Hori
authors.forename: R
authors.initials: R
authors.lastname: Matoba
authors.forename: T
authors.initials: T
authors.lastname: Niiyama
authors.forename: K
authors.initials: K
authors.lastname: Matsubara
mesh_terms: D001483:Base Sequence; D004247:DNA; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D004247:DNA