title: Direct detection of chicken genomic DNA for gender determination by thymine-DNA glycosylase.
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.
authors.affiliation: Department of Biological Sciences, University of Illinois at Chicago, Chicago, IL 60607, USA. nporat2@uic.edu
authors.forename: N
authors.initials: N
authors.lastname: Porat
authors.forename: K
authors.initials: K
authors.lastname: Bogdanov
authors.forename: A
authors.initials: A
authors.lastname: Danielli
authors.forename: A
authors.initials: A
authors.lastname: Arie
authors.forename: I
authors.initials: I
authors.lastname: Samina
authors.forename: A
authors.initials: A
authors.lastname: Hadani
mesh_terms: D000818:Animals; D030161:Avian Proteins / Q000235:genetics; D002645:Chickens / Q000235:genetics*; D004247:DNA / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D005819:Genetic Markers; D016678:Genome; D016133:Polymerase Chain Reaction; D012732:Sex Determination Analysis / Q000379:methods*; D044502:Thymine DNA Glycosylase / Q000032:analysis*
chemical_list: D030161:Avian Proteins; D004268:DNA-Binding Proteins; D005819:Genetic Markers; D004247:DNA; D044502:Thymine DNA Glycosylase