Screening for transgenic Japanese quail offspring.
After mosaic founder breeding pairs of Japanese quail start to produce fertile eggs, the hatchlings must be screened for germ-line transmission to the subsequent G1 generation. This article describes how to isolate hatchling genomic DNA from the chorioallantoic membrane (CAM), which remains inside the egg after hatching. Collecting genomic DNA from the CAM decreases the hatchling's stress during handling and eliminates the need for a blood draw. By following this protocol, the CAM of a single egg will provide 50 microg or more of high-quality genomic DNA. The article also describes how to screen the genomic DNA samples for the transgene by the polymerase chain reaction (PCR). PCR genotyping should be used for screening hatchlings with a nonfluorescent transgene or with a fluorescently labeled transgene that does not lend itself well to phenotypic screening.
Division of Biology and Biological Imaging Center, Beckman Institute, California Institute of Technology, Pasadena, CA 91125, USA.
Greg
G
Poynter
David
D
Huss
Rusty
R
Lansford
D000818:Animals; D030801:Animals, Genetically Modified; D049033:Chorioallantoic Membrane / Q000378:metabolism; D003370:Coturnix / Q000235:genetics*; D004247:DNA / Q000378:metabolism; D018014:Gene Transfer Techniques; D005821:Genetic Techniques; D016678:Genome; D005838:Genotype; D008856:Microscopy, Fluorescence / Q000379:methods; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods; D019076:Transgenes
D004247:DNA