A new approach to screen transgenic offspring using fluorescence in situ hybridization
- Lawrence Livermore National Laboratory, CA (United States)
- York Univ., Ontario (Canada)
In order to identify the presence of vector, specifically Lacl or LacZ, in putative transgenic mice rapidly and reliably, we developed a new method which utilizes fluorescence in situ hybridization (FISH). Transgenic mouse models are being used with increasing frequency for mutational and toxicological studies. By applying our method, an investigator can reliably determine the presence and the number of integration sites of a transgenic vector in numerous samples with less effort compared to conventional methods. This approach involves gazing a tail with a scalpel to obtain a blood smear on a microscope slide. After fixing the slide in 3:1 methanol: acetic acid, typical FISH analysis using biotinylated lambda DNA as the probe in performed. Our method eliminates the need for DNA extraction, blotting, and PCR, and yields results from a large number of individually identifiable cells from each animal. This assay is more accurate, reliable and easier to perform than conventional schemes presently used for screening transgenic animals. A particular advantage of this assay is the ability to discriminate between animals that are heterozygous and homozygous, something that eludes the PCR-based methods and Southern blotting. We have successfully analyzed over 95 samples with this method. Based on these results, we believe our system is more sensitive and accurate than conventional means of screening.
- DOE Contract Number:
- W-7405-ENG-48
- OSTI ID:
- 115151
- Report Number(s):
- CONF-9503160-; ISSN 0893-6692; CNN: Grant CA55861; TRN: 95:004640-0045
- Journal Information:
- Environmental and Molecular Mutagenesis, Vol. 25, Issue Suppl.25; Conference: 26. annual Environmental Mutagen Society meeting, St. Louis, MO (United States), 12-16 Mar 1995; Other Information: PBD: 1995
- Country of Publication:
- United States
- Language:
- English
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