2008•Zhongguo jiaqinRequires access
Establishment of Real-time Fluorescence PCR for GAPDH Gene of Chicken
SU Qingkang
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Abstract
According to the chicken GAPDH gene sequences available in GenBank,a pair of primers was designed for the establishment of a SYBR Green Ⅰ quantitative real-time PCR method for GAPDH gene of chicken. GAPDH was amplified by real-time fluorescence quantitative PCR from the plasmid DNA which was diluted to series standard concentrations,and the standard curves were established which indicate that there is a good linear function in statistics between the Ct value and the concentration gradient of standard plasmid DNA specimen. Under the established condition the reaction,the sensitivity of the methods was 10 copies/μL by the analysis of the dynamic curve. The recombinant plasmid and standard curve for GAPDH real-time quantitative PCR were constructed successfully. It provided the basis for GAPDH gene of chicken as a reference gene in quantitative analysis of mRNA expression.