Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5119

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Screening for Transgenic Japanese Quail Offspring

Greg Poynter, David Huss, and Rusty Lansford1

Division of Biology and the Biological Imaging Center, Beckman Institute, California Institute of Technology, Pasadena, CA 91125, USA

1Corresponding author (rusty{at}caltech.edu)

This article is also available in Emerging Model Organisms: A Laboratory Manual, Vol. 1. CSHL Press, Cold Spring Harbor, NY, USA, 2009.


INTRODUCTION

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 µg 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.


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This article has been cited by other articles:


Home page
 CSH ProtocolsHome page
G. Poynter, D. Huss, and R. Lansford
Japanese Quail: An Efficient Animal Model for the Production of Transgenic Avians
CSH Protocols, January 1, 2009; 2009(1): pdb.emo112 - pdb.emo112.
[Abstract] [Full Text]

Home page
 CSH ProtocolsHome page
G. Poynter, D. Huss, and R. Lansford
Generation of High-Titer Lentivirus for the Production of Transgenic Quail
CSH Protocols, January 1, 2009; 2009(1): pdb.prot5117 - pdb.prot5117.
[Abstract] [Full Text]

Home page
 CSH ProtocolsHome page
G. Poynter, D. Huss, and R. Lansford
Injection of Lentivirus into Stage-X Blastoderm for the Production of Transgenic Quail
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