TaqMan™ hPSC Scorecard™ 试剂盒，Fast 96 孔

No software update is required. Because the hPSC Scorecard Analysis Software is cloud based, you will always have access to the most up-to-date version of the software every time you log in.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center

The only way to view the Ct values in the current version is through the expression plot. We expect the software's Excel export to include the gene names and Ct values in a future version of the software.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center

A proprietary algorithm compares the Ct values for each marker set to the values in the reference database and calculates the score based on how well the expression correlates. In general, scores close to 0 indicate comparable expression to that of the reference standard using undifferentiated cells. Scores higher than 1 indicates up regulation relative to undifferentiated cells and less than -1 indicate down regulation relative to undifferentiated cells.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center

Thirteen pluripotent stem cell lines were used to generate the reference standard including:

• H9 ESC P28
• iPS BS3C P35
• iPS18C P29
• HUES9 P28
• HUES13 P56
• HUES28 P28
• HUES44 P25
• HUES48 P21
• HUES49 P21
• HUES53 P24
• HUES63 P46
• HUES64 P30
• HUES65 P29

The hPSC Scorecard Analysis Software performs a set of quality control checks upon data import to ensure the quality of the results.
You may see one of the following red flag warnings:

• Internal positive control: The expression levels of the housekeeping genes were low, signifying that there was an issue with the qRT-PCR. The sample is excluded from analysis and must be repeated.
• Bad Rox: The ROX dye is used as a passive reference during the qPCR. If the ROX dye is not detected, it is suggestive that some of the assay wells may not have received the proper amount of Master Mix. This may explain why a particular assay is not showing the anticipated gene expression level.
• Sendai Virus Detected (SEV): The hPSC Scorecard Panel includes a control to look for Sendai virus expression. A flag will appear if a Ct value of greater than 30 is observed for this control. On some rare occasions, the SEV oligonucleotides will cross react with other genes resulting in a false SEV flag in differentiated cells. If you see this flag, and did not anticipate Sendai virus detection, you may want to repeat the sample or look at the amplification plot (In the 96 well plate: well B9; in the 384 well plate: wells D3, D9, D15, and D21) to see if the automated Ct value call is real, or just an error.
• Insufficient Data: The analysis software requires a minimum number of genes to be amplified in each the four categories (i.e., self-renewal (“pluri”), ectoderm, mesoderm, and endoderm) to accurately analyze your data. If the minimum number of genes in each category does not amplify, or if data has been omitted by the user, the Insufficient Data flag will appear and the sample is excluded from analysis. The sample should be repeated.