Human Pluripotent Stem Cell Naïve State qPCR Array
For characterization of gene expression associated with hPSC naïve or primed states
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Overview
qPCR is used to determine changes in steady-state mRNA levels of gene expression across multiple samples, generally normalized to the relative expression of internal control genes. Gene-specific primers amplify target sequences within cDNA pools reverse-transcribed from mRNA and, as with TaqMan® technology, hybridized sequence-specific probes provide a fluorescent signal from the 5' exonuclease activity of Taq DNA polymerase. The accumulation rate of the fluorescent signal is used to quantify the sample cDNA, and thereby the amount of mRNA in the original cell lysate.
The Human Pluripotent Stem Cell (hPSC) Naïve State qPCR Array has validated primers and probes to detect 90 genes preselected for their demonstrated differential expression in naïve state and primed hPSCs and their derivatives, as well as 6 endogenous control genes. TBP (TATA box-binding protein) qPCR Control Template is provided as a synthetic DNA positive control. Naïve state hPSCs are self-renewing, retain the properties of the pre-implantation blastocyst, and can differentiate to both embryonic and extra-embryonic lineages. Primed hPSCs are also self-renewing, but they are restricted to differentiation along the three embryonic germ layers. Additional gene information and a qPCR analysis tool are available here.
Data Figures
Figure 1. Human Pluripotent Stem Cell Naïve State qPCR Array Plate Configuration
Figure 2.
Quantitative PCR analysis of gene expression of H1 hESCs cultured in a naive-like state compared to H1 hESCs cultured under a primed state. Genes associated with higher expression in a naive state than a primed state are shown in light green, genes expected to be more highly expressed in a primed state compared to a naive state are shown in orange and pluripotency associated genes expressed highly in both naive and primed states are shown in yellow. Genes associated with endoderm (red), mesoderm (light blue) ectoderm (dark green) and partially reprogrammed genes (dark blue) in feeder-free versus feeder-dependent tissue culture are also show. Y-axis measures the log change expression difference and X-axis indicates the name of the gene. Experiments were run on a Viia7 Fast (Applied Biosystems) Instrument and data analysis was performed using STEMCELL qPCR analysis program (www.stemcell.com/qPCRanalysis).
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Resources and Publications
Educational Materials (1)
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Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
