Quantitative analysis of trilineage potential of your PSCs
The Applied Biosystems TaqMan hPSC Scorecard Panel assesses pluripotency and trilineage differentiation potential using real-time qPCR assays and intuitive data analysis software. The hPSC Scorecard assay was developed in collaboration with Alex Meissner, and follows from his landmark publication (Cell 2011 144:439–452). The accompanying analysis software gives researchers anywhere in the world the ability to compare the gene expression profiles of their cell lines to a common reference set.
Why we chose the TaqMan hPSC Scorecard Assay
Hear from Jennifer Moore at RUCDR/Infinite Biologics on the benefits Scorecard has to offer.
Instrument requirements
See which instruments are compatible with the TaqMan hPSC Scorecard Panel and how to set them up.
Data analysis
The TaqMan hPSC Scorecard Panel includes access to free, intuitive web-based analysis software that allows researchers to quantitatively compare the gene expression profile of their sample to that of a reference set.
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The TaqMan hPSC Scorecard Panels quantitatively confirms pluripotency (trilineage differentiation potential)
for both embryonic and induced pluripotent stem cell lines. The hPSC Scorecard Panel (available in 96 and 384-well format) contains 94 predefined TaqMan Gene Expression assays (including endogenous controls) dried-down in assay plate wells. The panel of genes was developed in collaboration with Alex Meissner, and based on his landmark publication (Cell 144:439-452 (2011)).
The TaqMan hPSC Scorecard Panel provides:
- A quantitative assay for confirming pluripotency and trilineage differentiation potential
- Comparison of expression profiles to a reference standard
- An easy to use platform with pre-plated assays and dedicated, intuitive analysis software
Overview of protocol:
- Test undifferentiated, spontaneous differentiation (EB) or directed (monolayer) differentiation samples
- Extract and purify RNA
- Generate cDNA
- Run gene expression panel on a compatible qRT-PCR instrument
- Upload .eds files to the hPSC Scorecard Analysis Software
- Analyze data and export results from the convenient cloud-based analysis application
How to run the hPSC Scorecard Assay
Undifferentiated ESC/iPSC
Differentiated ESC/iPSC: Embryoid bodies (week 1) - or - Directly differentiated cells.
Sample requirements: >1 x 106 cells/sample
Undifferentiated ESC/iPSC
Feeder-dependent culture
Differentiated ESC/iPSC (EBs)
RNA Isolation
- TRIzol reagent
- RNase-free water
- Chloroform
- Isopropanol
- Ethanol
- DNA-free
Formulas
TRIzol organic phase extraction
Sample requirements: 1 mL TRIzol reagent per 6 cm culture dish
[RNA] (mg/mL) = A260 * 40
Recommended A260/A280 ratio = 1.6 – 1.8
8 reverse transcription (RT) reactions per RNA sample
Experiment type: Standard Curve
Analyze data from the TaqMan hPSC Scorecard Panel using the intuitive analysis software
Sign in to the hPSC Scorecard Analysis Software
- Learn more about software and data analysis
Undifferentiated ESC/iPSC
Differentiated ESC/iPSC: Embryoid bodies (week 1) - or - Directly differentiated cells.
Sample requirements: >1 x 106 cells/sample
Undifferentiated ESC/iPSC
Feeder-dependent culture
Differentiated ESC/iPSC (EBs)
RNA Isolation
- TRIzol reagent
- RNase-free water
- Chloroform
- Isopropanol
- Ethanol
- DNA-free
Formulas
TRIzol organic phase extraction
Sample requirements: 1 mL TRIzol reagent per 6 cm culture dish
[RNA] (mg/mL) = A260 * 40
Recommended A260/A280 ratio = 1.6 – 1.8
8 reverse transcription (RT) reactions per RNA sample
Experiment type: Standard Curve
Analyze data from the TaqMan hPSC Scorecard Panel using the intuitive analysis software
Sign in to the hPSC Scorecard Analysis Software
- Learn more about software and data analysis
How to Prepare the Applied Biosystems™ TaqMan® hPSC Scorecard™ Panel
For Research Use Only. Not for use in diagnostic procedures.