何首乌实时荧光定量PCR内参基因筛选

为筛选适宜何首乌Polygonum multiflorum不同组织基因表达分析的内参基因,从何首乌转录组数据中选取Actin,GAP-DH,SAND,PP2A,TIP41常用的5个候选内参基因,设计特异性引物.通过实时荧光定量PCR,利用geNorm,NormFinder,BestKesper,Delta CT以及Re...

Reference gene screening for Real-time quantitative PCR in Polygonum multiflorum

To select suitable references gene of Polygonum multiflorum for gene expression analysis in different tissues,five candidate reference genes like Actin,GAPDH,SAND,PP2A,TIP41 were selected from the transcriptome data of P.multiflorum,then the specific primers were designed.The expression stability of the five reference genes in different tissues of P.multiflorum was analyzed by Real-time quantitative PCR through avilable analysis methods such as geNorm,NormFinder,BestKeeper,Delta CT and RefFinder,to ensure the reliability of the analysis results.The results showed that there were significant differences in the expression levels and stability of candidate genes in different tissues of P.multiflorum.Ct distribution analysis of the expression levels of candidate genes showed that the expression levels of Actin and GAPDH genes were relatively high in different tissues,while the expression levels of SAND,PP2A and TIP41 were lower.The stability of each candidate gene was analyzed by different methods.The results of geNorm analysis showed that the expression of PP2A and GAPDH was the most stable,the expression stability of SAND was the worst,the stability of PP2A was the highest in both NormFinder and Delta CT,the stability of SAND was the lowest,and the stability of Actin was the most stable in BestKeeper analysis.Through the comprehensive evaluation and analysis of the stability of candidate genes by RefFinder,it is con-cluded that the stability of PP2A gene is the highest,followed by GAPDH,Actin,TIP41,SAND,and SAND gene is the worst.Therefore,the PP2A gene is an ideal reference gene for the analysis of gene expression in different tissues of P.multiflorum.