The mapping of transgenes by fluorescence in situ hybridization on G-banded mouse chromosomes.
A highly sensitive method for the mapping of transgenes and other genes in the mouse genome is described. This technique combines high-resolution G-banding and fluorescence in situ hybridization (FISH) with either biotin/avidin-FITC or digoxigenin-anti-digoxigenin-FITC, the latter being the more sensitive. Banding patterns are obtained with trypsin/Giemsa-treated slides, and sensitivity is greatly increased by the use of mouse Cot-1 DNA. With this protocol, four different 14.5-kb human Cu/Zn-superoxide dismutase transgene insertions ranging in copy number from 2 to 8 have been localized to four different mouse chromosomes. The utility and sensitivity of this procedure were verified with a Chromosome (Chr) 16-specific cosmid probe, H22, as well as with the mapping of a high-copy-number human beta-amyloid/A4 transgene.
Shi
Y P
YP
Department of Pediatrics, University of California, San Francisco 94143.
Huang
T T
TT
Carlson
E J
EJ
Epstein
C J
CJ
D000818:Animals; D002871:Chromosome Banding / Q000379:methods; D002874:Chromosome Mapping / Q000379:methods*; D003360:Cosmids; D015342:DNA Probes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D013482:Superoxide Dismutase / Q000235:genetics
D015342:DNA Probes; D013482:Superoxide Dismutase