An evaluation of CHD‐Specific primer sets for sex typing of birds from feathers

Abstract

The amplification of the highly conserved chromo-helicase-DNA binding region found in both the Z and W chromosome was evaluated with three sets of primers (P8/P2, 1237L/1272H and 2550F/2718R). DNA extracted from feathers through a simple boiling method was used to address its reliability in generating the sex-linked bands. All the bird samples, including the seven bird families that have not been reported previously, were successfully amplified with the primer set 2550F/2718R. The resulting polymerase chain reaction products showed clearly resolved fragments on a conventional agarose gel electrophoresis with size differences ranging from 80 to 540 bp between the two respective ZW gene copies. Although the P8/P2 primer was not as effective under the same conditions, it was able to produce well-resolved Z and W bands from bird species under the Antidea family, whereas the 2250F/2718R primer set only produced a single amplified fragment of a different size between the male and the female. Zoo Biol 27:62-69, 2008. (c) 2007 Wiley-Liss, Inc.

Molecular sex determination of 20 bird species protected in the Republic of Serbia

“…In male birds only one band is visualized at approximately 650 bp (CHD-Z). These intron sizes are in accordance with previously published data (Ong and Vellayan, 2008). DISCUSSION The method used in this study gave good results with species belonging to orders Podicipediformes, Ciconiiformes, Anseriformes, Falconiformes, Galliformes, Columbiformes, Passeriformes, Charadriiformes including six species (Anser anser, A. fabalis, Buteo buteo, Coturnix coturnix, Perdix perdix, and Scolopax rusticola) with no previous data available on molecular sexing.…”

“…DISCUSSION The method used in this study gave good results with species belonging to orders Podicipediformes, Ciconiiformes, Anseriformes, Falconiformes, Galliformes, Columbiformes, Passeriformes, Charadriiformes including six species (Anser anser, A. fabalis, Buteo buteo, Coturnix coturnix, Perdix perdix, and Scolopax rusticola) with no previous data available on molecular sexing. In Platalea and also Accipitridae and Anatidae species previous attempts at molecular sexing gave ambiguous results (Wang et al, 2007;Ong and Vellayan, 2008). Also a larger number of samples was included for species where the protocol by Vucicevic et al (2012) was previously employed, confirming the reliability and robustness of the method.…”

Application of Two Molecular Sexing Methods for Indonesian Bird Species: Implication for Captive Breeding Programs in Indonesia

“…A problem of thick band was never found with 2550F/2718R primers, so incorrect identification of female to male bird was avoided. The results of amplified bands (CHD-W and CHD-Z) produced in this study were the same as previous results obtained by Ong and Vellayan (2008);Vucicevic et al (2012b), i.e. in female birds two amplified bands is visualized around 400 and 650 bp (CHD-W and CHD-Z) and in male birds only one band is visualized at approximately 650 bp (CHD-Z).…”

Short Communication Non-invasive sex identification of the white-bellied sea eagle (Haliaeetus leucogaster) through genetic analysis of feathers

“…In this study, the use of feathers as a source of DNA has been proven as an efficient and effective technique for genetic sexing of the white-bellied sea eagle. This technique has also been applied in other Falconiformes species (see e.g., Sacchi et al, 2004; Rudnick et al, 2007;Ong and Vellayan, 2008). Because avian blood cells are nucleated, the total amount of DNA extracted from feathers seems to be sufficient for conducting this research (Ong and Vellayan, 2008).…”

“…The use of the P2/P3 primer combination was suitable for sex discrimination of the white-bellied sea eagle (Figure 1) and 38 other species of raptors (representing four families), which was reported by Norris-Caneda and Elliot (1998). The primer used also provided a high confidence level for establishing the sex of birds even without the use of polyacrylamide gels as required in some bird species (Griffiths et al, 1998;Ong and Vellayan, 2008).…”