Nadine Nassif1, Nathan Greve1, Soshana Svendsen1, Clive Svendsen2 and Douglas R. Storts1
1Promega Corporation, Madison, WI
2University of Wisconsin-Madison
Stem cell differentiation is routinely monitored by measuring gene expression using real-time reverse transcription PCR. The Plexor® Differential Expression System is a novel, real-time PCR system for the detection and relative quantitation of RNA expression levels, and can be used to monitor stem cell differentiation using multiplexed amplification.
Optimized primer pairs allow users to quantitatively amplify specific transcripts in a two-color duplex reaction. This duplex amplification strategy enables the user to amplify a transcript of interest as well as a reference transcript (GAPDH) in a single reaction.
The first set of optimized primer pairs to be made available will focus on transcripts associated with stem cell pluripotency: NANOG, SOX2, OCT3/4, LIN28, KLF4, and cMYC, each in a duplex with GAPDH. These primers target genes that are expressed at high levels in pluripotent cells (i.e., stem cells and induced pluripotent stem cells [iPS]). In contrast, these genes are expressed at low levels in differentiated cells.