Thermo Fisher Scientific
QuantStudio 5 Real-Time PCR System, 384-well
A28140
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I'm interested in understanding the molecular mechanism of hedgehog signal transduction. One of the metrics that I use for assessing the hedgehog signaling pathway output is measurement of Gli1 mRNA levels. Gli1 is an immediate hedgehog ligand induced target gene. Quantitative PCR (qPCR) for Gli1 was performed using QuantStudio 5 Real-Time PCR System. This equipment was purchased after passing rigorous testing phase during demo.
qPCR
cDNA from NIH-3T3 cells
Confluent NIH-3T3 cells were serum starved and left untreated or treated with hedgehog ligand (sonic hedgehog) for 4 hours. RNA was extracted and cDNA was synthesized using standard procedures. Gli1 mRNA levels were measured by qPCR (normalized to a house keeping gene).
Follow manufacturer's instructions!
Gli1 mRNA levels were found to be elevated by ~30-40 with hedgehog ligand treatment compared to untreated cells. This instrument is very easy to use and results were found to be highly reproducible.
Easy to use and occupies less space
N/A
Easy to use
None
Easy to use and since this is a LED based system, the capital cost is relatively cheaper compared to laser based systems.