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Record number: 1, Textual support number: 1 CONT

Two strategies have been used to clone specific sequences of eukaryotic, genomic DNA in bacteriophage lambda vectors. The first involved cloning of DNA preparations that had been highly enriched for the sequences of interest. Total genomic DNA was digested to completion with a restriction endonuclease and the resulting fragments were separated by preparative gel electrophoresis, reverse-phase column chromatography, or both. The fractions containing the DNA sequences of interest were identified by hybridization to appropriate probes and cloned in bacteriophage lambda vectors. Because these fractionation procedures may yield an overall enrichment of as much as 100-fold to 300-fold, the number of recombinant bacteriophages that had to be generated and screened for the DNA sequences of interest could be significantly reduced. Subsequently, the development of efficient in vitro packaging systems and in situ hybridization techniques eliminated the need to enrich the starting DNA for the sequences of interest.  1, fiche 1, Anglais, - reverse%20phase%20column%20chromatography

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