How can I improve curve dynamics in qPCR?

03 November 2022 1 8K Report

I am working on a qPCR with specific probes, however my standard titration does not display any titration down. The amplification plot shows high intensity of every point (including zero) with a Ct around 8-12 cycles. The amplification plot looks the same with SYBR green and probe system master mix. Not considering pipetting, centrifugation, nor mixing, does anyone have any suggestions?

Amy Klocko

Maybe the machine needs cleaning/calibrating? You could try loading a plate of just water, or water and dye in some wells and see how that reads as a control.

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