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Increment of efficiency in the identification of noble genes by colony hybridization assay. (Screening of low redundant clones from human fetal cDNA library).
Kim, J W; Lee, I A; Lee, Y H; Song, J C; Choe, Y K; Hahn, Y S; Chung, J H; Chung, T W; Choe, I S.
Afiliación
  • Kim JW; Molecular and Cellular Biology Division, Korea Research Institute of Bioscience and Biotechnology, KIST, Taejon, Korea.
Biochem Mol Biol Int ; 44(2): 225-33, 1998 Feb.
Article en En | MEDLINE | ID: mdl-9530506
For the rapid identification of noble genes in a specific tissue by computer analysis from the cDNA sequences determined by single-pass cDNA sequencing, clone redundancy was one of the major obstacles. To facilitate the efficiency in identification of noble genes, it was necessary to reduce the number of clones to be sequenced by eliminating the redundant clones for a rapid analysis. In order to increase the probability of isolating noble sequences from the cDNA clones of human fetal liver tissue origin, colony hybridization assay was adopted and redundant clones were efficiently removed. Four cDNA clones highly redundant in the human fetal liver cDNA libraries including alpha-globin, gamma-globin, serum albumin and H19 RNA sequences were selected as the probes. Two hundreds and sixty two cDNA clones were randomly selected and tested with the probes for hybridization properties. The identity of each cDNA clone giving positive or negative signals in the hybridization assay was determined by DNA homology search with the nucleic acid databases. Among the 76 clones giving positive signals, 57 clones (75%) were found to be identical to the probe sequences and could be eliminated by colony hybridization assay before neucleotide sequencing.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Sonda Molecular / Clonación Molecular / ADN Complementario Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Humans Idioma: En Revista: Biochem Mol Biol Int Asunto de la revista: BIOLOGIA MOLECULAR / BIOQUIMICA Año: 1998 Tipo del documento: Article Pais de publicación: Reino Unido
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Sonda Molecular / Clonación Molecular / ADN Complementario Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Humans Idioma: En Revista: Biochem Mol Biol Int Asunto de la revista: BIOLOGIA MOLECULAR / BIOQUIMICA Año: 1998 Tipo del documento: Article Pais de publicación: Reino Unido