The invention relates to primers, kit and method to detect transcriptional level of
purine nucleotide phosphorylase PNP
gene in
macaque liver by RT-qPCR (real-time quantitative
polymerase chain reaction), and belongs to the technical field of
molecular biology. A template is made with cDNA synthesized by reverse transcription of
total RNA extracted from fresh
macaque liver tissue; a PCR primer combination is used to perform real-time fluorescent quantitative PCR amplification; Ct values of PNP
gene fragments and
reference gene GAPDH fragments,
dissolution peaks, and amplification efficiency are attained; multiple-expressed
delta delta C(t) value of the homogenize PNP
gene is acquired by
conventional treatment; relative expression value of the transcriptional level of PNP gene is acquired finally. An effective means to study PNP functionality of
macaque and influence of relevant drugs upon PNP is provided; in addition, the primers, kit and method herein are applicable to real-time quantitative PCR detection and have the advantages of good operational simplicity, high
repeatability, low detection cost, high sensitivity, high specificity and good convenience of popularization and application.