Establishment of fluorescent quantitative real time RT-PCR method for detection of the expression of nerve growth factor mRNA

Objective:To establish a new method for detecting the expression of nerve growth factor messenger ribonucleic acid(NGF mRNA) in cerebrum tissue of rats.Methods:Total RNA was isolated from cerebrum tissue by Trizol-one-step method.Specific oligonucleotide of NGF mRNA gene fragments were amplificated by RT-pCR.The method of detection of NGF mRNA expression using SYBR Green Ⅰ real time RT-pCR was constructed.The specificity of the method was confirmed by melting curve.Housekeeping gene -actin was applied to rectify the quantity of NGF.Results:SYBR Green real-time pCR was developed with high specificity and sensitivity for detecting of rat NGF mRNA.Conclusions:The method of detection of NGF mRNA expression using real time RT-pCR is constructed successfully.