Identification of a tumor marker chromosome by flow sorting, DNA amplification in vitro, and in situ hybridization of the amplified product
- Univ. of Amsterdam (Netherlands)
- State Univ. of Groningen (Netherlands)
A method combining flow sorting and molecular cytogenetic techniques was used to identify an unknown marker chromosome in the bladder tumor cell line J82. The marker chromosome was isolated by dual parameter sorting after staining with Hoechst 33258 and chromomycin [Lambda]3. DNA amplification of 300 isolated chromosomes by polymerase chain reaction using the Alu-primer Bk33 and the Lines-primer LH5 was carried out. The DNA was labelled using Bio-11-dUTP and applied to human lymphocyte metaphase cells in a suppressive in situ hybridization procedure. Fluorescence was visible over chromosome 20 and over the distal one-half of 6p. Together the fluorescent regions accounted for only 60% of the marker length, indicating a possible duplication of chromosome 20 material. This was subsequently confirmed by applying bicolor in-situ hybridization using chromosome 6- and 20-specific DNA libraries to metaphase cells of the J82 culture.
- OSTI ID:
- 6827863
- Report Number(s):
- CONF-9303114--
- Journal Information:
- Cytometry (Baltimore); (United States), Journal Name: Cytometry (Baltimore); (United States) Vol. 6; ISSN CYTODQ; ISSN 0196-4763
- Country of Publication:
- United States
- Language:
- English
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