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Abstract
1. Birds, especially nestlings, are generally difficult to sex by morphology and early detection of chick gender in ovo in the hatchery would facilitate removal of unwanted chicks and diminish welfare objections regarding culling after hatch.
2. We describe a method to determine chicken gender without the need for PCR via use of Thymine-DNA Glycosylase (TDG). TDG restores thymine (T)/guanine (G) mismatches to cytosine (C)/G. We show here, that like DNA Polymerase, TDG can recognise, bind and function on a primer hybridised to chicken genomic DNA.
3. The primer contained a T to mismatch a G in a chicken genomic template and the T/G was cleaved with high fidelity by TDG. Thus, the chicken genomic DNA can be identified without PCR amplification via direct and linear detection. Sensitivity was increased using gender specific sequences from the chicken genome.
4. Currently, these are laboratory results, but we anticipate that further development will allow this method to be used in non-laboratory settings, where PCR cannot be employed.
Acknowledgements
This work is dedicated to the memory of Professor Aryeh Hadani, who served as the head of the Kimron Veterinary Institute (K.V.I) between 1963–1976. We thank M. Kamensky, I. Alchanati, O. Tsioni and the researchers at QBI Enterprises Ltd. for helpful discussions. We thank Prof. Kay for review of the manuscript (University of Illinois at Chicago). We thank the Ishaya Horowitz fund, NiKagen Ltd., and K.V.I for their support. Patent application no. 12/021,307.