Sex ratio determination in bovine semen: A new approach by quantitative real time PCR

Parati, Katia; Bongioni, G.; Aleandri, R.; Galli, Andrea

doi:10.1016/j.theriogenology.2006.07.007

Cited by 65 publications

(50 citation statements)

References 24 publications

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“…In general, these methodologies were developed mainly based on amelogenin gene (AMELX and AMELY) (Ennis and Gallagher, 1994; Chen et al, 1999; Fontanesi et al, 2008), zinc finger gene (ZFX and ZFY) (Aasen and Medrano, 1990; Kirkpatrick and Monson, 1993; Zinovieva et al, 1995; Lockley et al, 1997) and the sex determination region of the Y chromosomal gene (SRY) (Fu et al, 2007; Lu et al, 2007; Shi et al, 2008) Alternatively, the bovine-specific repetitive sequence BRY-1 (Matthews, 1990) and the single copy sequence BOV97 M (Miller and Koopman, 1990) are also used in the analysis for the sexing of cattle. More recently, based on real time PCR both SYBR Green (Ballin and Madsen, 2007) and TaqMan (Parati et al, 2006) for bovine sex determination have also been reported.…”

Section: Introductionmentioning

confidence: 99%

Sex Determination of Cattle Meat by Polymerase Chain Reaction Amplification of the DEAD Box Protein (DDX3X/DDX3Y) Gene

Gokulakrishnan

Kumar

Sharma

et al. 2012

Asian Australas. J. Anim. Sci

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Determination of sex origin of cattle meat by fast and reliable molecular methods is an important measure to ensure correct allocation of export refunds particularly in European countries and also female cattle (cow) slaughter is legally banned in India because of religious beliefs. Based on the DEAD box protein gene located on the X and Y chromosomes, 2 pair of primers were designed and the system of PCR was optimized. Upon PCR amplification, male tissue showed 2 bands, while female tissue resulted in only one band. The accuracy and specificity of the primers was assessed using DNA template extracted from cattle meat of known sex. The protocol was subjected to a blind test and showed 100% concordance, proving its accuracy and reliability.

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Section: Introductionmentioning

confidence: 99%

Sex Determination of Cattle Meat by Polymerase Chain Reaction Amplification of the DEAD Box Protein (DDX3X/DDX3Y) Gene

Gokulakrishnan

Kumar

Sharma

et al. 2012

Asian Australas. J. Anim. Sci

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“…However, OptiPrep was not effective to separate X sperm. The qPCR has recently been used as a reliable technique for the quantification of X and Y sperm cells in semen samples, especially to validate techniques for sexing sperm (Joerg et al, 2004;Parati et al, 2006;Puglisi et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, it was possible to quantify the proportion of X and Y sperm in semen samples before centrifugation without any adjustments by using linear regression, as described in other studies (Parati et al, 2006;Puglisi et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

“…DNA samples were subjected to electrophoresis in 2% agarose gel for visualization of genomic DNA. The qPCR procedure (Parati et al, 2006) was used to determine the enrichment of samples with X sperm, after centrifugation in density gradients, without the need to perform validation by means of in vitro production of embryos and subsequent sexing by conventional PCR. For internal control a negative control with no DNA added was used, a control with DNA extracted from the blood of a male bovine and with DNA extracted from spermatozoa not centrifuged in density gradients.…”

Section: Methodsmentioning

confidence: 99%

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Comparative validation using quantitative real-time PCR (qPCR) and conventional PCR of bovine semen centrifuged in continuous density gradient

Resende

Lucio

Perini

et al. 2011

Arq. Bras. Med. Vet. Zootec.

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Comparative validation using quantitative real-time PCR (qPCR) and conventional PCR of bovine semen centrifuged in continuous density gradient [Validação comparativa utilizando PCR quantitativo em tempo real (qPCR) ABSTRACTThe objective of the present study was to determine the sperm enrichment with X-bearing spermatozoa, after one centrifugation in a Percoll or OptiPrep continuous density gradient, using quantitative real-time polymerase chain reaction (qPCR) of sperm DNA and resultant in vitro-produced bovine embryos by PCR. Frozen/thawed sperm was layered on density gradients and the tubes were centrifuged. Supernatants were gently aspirated and the sperm recovered from the bottom of the tubes. Cleavage and blastocyst rates were determined through in vitro production of embryos and PCR was performed to identify the embryos' genetic sex. A difference in blastocyst rate was found in the Percoll treatment compared to OptiPrep (P<0.05). The percentage of female embryos in the Percoll and OptiPrep groups was 62.0% and 47.1%, respectively. These results were confirmed by qPCR of spermatozoa DNA and underestimation was seen only in the Percoll group. It was possible to sexing sperm using simple approach.

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“…Alternatively, the bovine-specific repetitive sequence BRY-1 (Matthews & Reed, 1992) and the single copy sequence BOV97 M (Miller & Koopman, 1990) are also used in the analysis for the sexing of bovine. More recently, based on real time PCR, both SYBR Green (Ballin & Madsen, 2007) and TaqMan (Parati, Bongioni, Aleandri, & Galli, 2006) technology for bovine sex determination have been reported. Moreover, studies have demonstrated that SRY gene can also be used as a target DNA for embryos sexing by PCR amplification (Fu et al, 2007;Shi, Yue, Ren, Lei, & Zhao, 2008).…”

Section: Introductionmentioning

confidence: 99%

A PCR assay for sex determination of yak (Bos grunniens) meat by amplification of the male-specific SRY gene

et al. 2010

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Sex ratio determination in bovine semen: A new approach by quantitative real time PCR

Cited by 65 publications

References 24 publications

Sex Determination of Cattle Meat by Polymerase Chain Reaction Amplification of the DEAD Box Protein (DDX3X/DDX3Y) Gene

Sex Determination of Cattle Meat by Polymerase Chain Reaction Amplification of the DEAD Box Protein (DDX3X/DDX3Y) Gene

Comparative validation using quantitative real-time PCR (qPCR) and conventional PCR of bovine semen centrifuged in continuous density gradient

A PCR assay for sex determination of yak (Bos grunniens) meat by amplification of the male-specific SRY gene

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