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2007•China Animal Husbandry & Veterinary MedicineRequires access

Establishment of Real-time PCR for β-actin Gene of Chicken

Hua Yue

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Abstract

According to the chicken β-actin gene sequences available in GenBank,a pair of primers was designed for establish a SYBR Green I quantitative real-time PCR method for β-actin gene of chicken.To establish the standard curve,the product of conventional PCR served as a standard. The analysis of melting curve was also carried out.The results show the linear range of Ct value was from 12 to 31 with a good correlation coefficient(r=0.996).The melting curve show a single peak with a 88±0℃ Tm value.The real-time PCR assay developed in this study can detect β-actin in expand range with high efficiency and less time.The assay provide the basis for β-actin gene of chicken as a reference gene in quantitative analysis of mRNA expression.

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What this paper is about

According to the chicken β-actin gene sequences available in GenBank,a pair of primers was designed for establish a SYBR Green I quantitative real-time PCR method for β-actin gene of chicken.To establish the standard curve,the product of conventional PCR served as a standard. The analysis of melting curve was also carried out.The results show the linear range of Ct value was from 12 to 31 with a good correlation coefficient(r=0.996).The melting curve show a single peak with a 88±0℃ Tm value.The real-time PCR assay developed in this study can detect β-actin in expand range with high efficiency and less time.The assay provide the basis for β-actin gene of chicken as a reference gene in quantitative analysis of mRNA expression.

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Available abstract

According to the chicken β-actin gene sequences available in GenBank,a pair of primers was designed for establish a SYBR Green I quantitative real-time PCR method for β-actin gene of chicken.To establish the standard curve,the product of conventional PCR served as a standard. The analysis of melting curve was also carried out.The results show the linear range of Ct value was from 12 to 31 with a good correlation coefficient(r=0.996).The melting curve show a single peak with a 88±0℃ Tm value.The real-time PCR assay developed in this study can detect β-actin in expand range with high efficiency and less time.The assay provide the basis for β-actin gene of chicken as a reference gene in quantitative analysis of mRNA expression.

Key concepts: Melting curve analysis, GenBank, Real-time polymerase chain reaction, Standard curve, SYBR Green I, Gene, Molecular biology, Biology

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