Pushing gene expression throughput
PCR has become a common method for monitoring cellular gene expression, but when it comes to examining the impact of large numbers of drug candidates on cells, PCR suffers from throughput limitations.
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OSAKA, Japan—PCR has become a common method for monitoring cellular gene expression, but when it comes to examining the impact of large numbers of drug candidates on cells, PCR suffers from throughput limitations. In Toxicology in Vitro, however, scientists at Takeda Pharmaceuticals looked to ramp up the throughput of their PCR-based assay by modifying it to run in the multiwell nuclease protection-based ArrayPlate qNPA.
The researchers used both real-time PCR and ArrayPlate to monitor the impact of 80 compounds on the expression of 12 genes associated with phospholipidosis in a liver carcinoma cell line. They noted that the ArrayPlate offered a linear response over a range of starting concentrations and that well-to-well, plate-to-plate and day-by-day repeatability was good. They also noted that the phospholipidosis scores between ArrayPlate and real-time PCR correlated significantly (R2=0.95).
"Because it is 96-well microplate based and does not require RNA extraction, reverse transcription or gene amplification, the ArrayPlate not only provided greater day-to-day and sample-to-sample repeatability but also significantly increased testing throughput and cost-efficiency," the authors concluded.