qPCR 用cDNA参照
品牌	Code No.	包装量	价格
Clontech	636692	25 Rxns	952 元
Clontech	636693	100 Rxns	2,845 元
Clontech	639653	25 Rxns	952 元
Clontech	639654	100 Rxns	2,845 元

Reference cDNA for Real-Time qPCR

Measurements of mRNA expression levels—whether by Northern analysis, ribonuclease protection, or real-time quantitative PCR—are usually standardized by comparing the data to that obtained for an internal or endogenous reference gene. Housekeeping genes such as beta-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are most often used because their expression levels are expected to remain constant under different treatment conditions. Unfortunately, this assumption is not always valid, and results based on housekeeping genes alone can be biased (1). A better method is to normalize your data using our qPCR Human Reference cDNA, the only total cDNA control derived entirely from human tissues (2).

qPCR Human Reference cDNA is the ideal control for comparing data from different quantitative PCR (qPCR) experiments. Because it is prepared from a total RNA pool collected from several different tissues, our qPCR Human Reference cDNA provides broad gene coverage, as shown by microarray analysis of the RNA starting material. RNA, and therefore cDNA, prepared from whole tissues provides better gene representation with less variation than RNA made from cell lines (data not shown). Moreover, PCR analysis shows that our Total RNA is virtually free of genomic DNA (3). This allows for a more accurate measurement of transcript copy number. Both high and low abundance genes are well represented allowing preparation of a wide range of serially diluted standards for each qPCR assay. Lot-to-lot variation of Reference cDNA is minimal because the RNA source is prepared on an industrial scale.

Product image

High-, medium- and low-abundance gene targets are easily detected in qPCR Human Reference cDNA. Oligo dT-primed (Panels A–C) and random oligo-primed (Panels D–F) qPCR Human Reference cDNA samples were each serially diluted fivefold such that the final quantities of cDNA template in the qPCR reactions were 20 ng, 4 ng, 800 pg, 160 pg, 32 pg,and 6.4 pg. Data were obtained on a Stratagene Mx3000P real-time PCR instrument. The primer sets for all three genes demonstrate the ability to detect a wide range of signals from the serially diluted samples. PPIA = peptidylprolyl isomerase A (cyclophilin A; high-abundance); HPRT = hypoxanthine phosphoribosyltransferase I (medium-abundance); PBGD = porphobilinogen deaminase (low-abundance). Slope and R2 values refer to the line determinedby plotting Ct values versus template quantity.