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Construction and characterization of a DNA library from mouse chromosomes 19 purified by flow cytometry
Author(s) -
Baron Bruno,
Métézeau Philippe,
KieferGachelin Hélène,
Goldberg Michel E.
Publication year - 1990
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/0248-4900(90)90322-t
Subject(s) - biology , chromosome , microbiology and biotechnology , karyotype , dna , genetics , genomic library , genome , flow cytometry , recombinant dna , southern blot , gene , base sequence
Summary— In spite of the constant development concerning physical mapping of eukaryotic genomes, the mouse chromosome 19 remains poorly characterized. In order to improve the possibilities for studying this chromosome, we have constructed a chromosome‐specific Eco RI DNA fragment library from mouse chromosomes 19 sorted by flow cytometry. The resulting library contains about 3 × 10 4 recombinant clones. The identified inserts range in size from about 0.2–10 kb, with a 4 kb average size and with no observable redundancy. The purity of the library has been analyzed by flow‐blot. For that purpose, chromosomes from 2 cell lines, 1 with a normal karyotype and 1 with translocated chromosome 19, were sorted on nylon filters and hybridized with 9 clones of the library. Results show that 5 clones out of the 9 clearly originate from sorted chromosomes 19 and 3 and are likely to be derived from its DNA, thus indicating that the library of chromosome 19 is of high purity.