Hi, I read through your paper. Interesting method.
In figure 1A, the workflow includes a standard PCR step before Sanger. Workflow-wise, wouldn't it still have the same bottleneck as qPCR test, i.e. limited by 96/384-well instrument runs?
Thanks; that is a good question. In our laboratory, we have a ratio of 10:1 for PCR to qPCR instruments. In the new laboratory that we are constructing, the ratio is 50:1. It was similar at Stanford academic laboratories during my PhD. Standard PCR instruments are inexpensive and very common. qPCR instruments are definitely not as common, as they are very specialized instruments for a few use-cases.
Most clinical laboratories would have 10 to 50 PCR instruments that they can use to run the initial amplification reaction in parallel before Sanger sequencing. Also, Sanger sequencing uses a plate feeder, so you can add new plates on top as the second round of PCR reactions finish.
But, more importantly, the qSanger can by-pass RNA extraction, which seems to be an important bottleneck in the RT-qPCR workflow.
In figure 1A, the workflow includes a standard PCR step before Sanger. Workflow-wise, wouldn't it still have the same bottleneck as qPCR test, i.e. limited by 96/384-well instrument runs?