StemElite™ Gene Expression System Technical Manual

Literature # TM323

The StemElite™ Gene Expression System is a novel real-time quantitative PCR (qPCR) system for the detection and relative quantification of RNA expression levels associated with the differentiation state or “potency” of cells. This technical manual describes the StemElite™ Gene Expression System and provides instructions for use, including reaction setup and data analysis using the Plexor® Analysis Software.
The Plexor® technology takes advantage of the specific interaction between two modified nucleotides to achieve quantitative PCR analysis. One of the PCR primers contains a modified nucleotide (iso-dC) linked to a fluorescent label at the 5´ end. The second PCR primer is unlabeled. The reaction mix includes deoxynucleotides and iso-dGTP modified with the quencher dabcyl. Dabcyliso- dGTP is incorporated opposite the iso-dC residue in the primer. The incorporation of the dabcyl-iso-dGTP at this position results in quenching of the fluorescent dye on the complementary strand and a reduction in fluorescence, which allows quantification during amplification.