How to Use the TaqMan hPSC Scorecard Panel
The TaqMan hPSC Scorecard Panel is a qPCR-based assay that measures expression of pluripotency in germ layer-specific markers. With this set of genes, you can accurately evaluate your pluripotent stem cells. This video will show you how to prepare and load your hPSC Scorecard plate. To get started, you can use human embryonic stem cells or human induced pluripotent stem cells, iPSCs, that are growing on feeder or in feeder-free conditions. You can also run this assay on embryoid bodies. The panel will accurately predict lineage bias from samples taken as early as day 5. The qPCR reaction for the hPSC Scorecard Panel can be run either on a 96-well plate or a 384-well plate. The 96-well plate can fit one sample, and the 384-well plate can fit 4 samples. To set up the qRT-PCR reactions, simply add TaqMan Gene Expression Master Mix to your prepared cDNA. We recommend setting up your reverse transcription reactions of one sample per 8-tube PCR strip to make this step more convenient. Then, load 8 µL per well into the hPSC Scorecard plate using a multichannel pipette. For 96-well plates, one well of the cDNA strip is sufficient to load one row of 12 in the qPCR plate. For 384-well plates, one well of the cDNA strip is sufficient to load 2 rows of 6 in the qPCR plate. When the pipetting is complete, seal the plate with an optical cover and centrifuge the plate. Place the plate in the qPCR instrument, then load the template file, and run the qPCR. A Thermo Fisher Scientific account must be created in order to use the hPSC Scorecard software. After the qPCR is complete, save the files to be used in the TaqMan hPSC Scorecard analysis software. With this powerful data analysis tool, the TaqMan hPSC Scorecard Panel enables accurate confirmation of the pluripotency and verification of trilineage differentiation potential.