Question about large-scale PCR

Hi everyone, I was approached by a colleague that was interested in automating their PCR workflow for amplicon NGS. Currently, their workflow involves setting up the PCR reaction in a 15 mL tube, splitting the PCR reaction into all the wells of a 96-well plate, and pooling the reactions back after thermocycling.

The straightforward way to automate this process is having a liquid handler doing the reaction set up and post-amplification pooling with an ODTC. I was wondering if anyone else had encountered a more fit-for-purpose solution or can think of a different way to handle and automate large volume thermocycling.

Not in NGS so take this with a grain of salt, but for large quantities of DNA I’d start thinking about isothermal amplification like RCA. I’ve taken amplicons, used KLD to circularize, then RCA’d for gross amplification. I then restriction digested to return to linear fragments.

Otherwise? I’ve read some cool stuff about continuous flow microfluidic thermocyclers. Nothing commercial, though.

Thanks, I’ll give a bit more thought to how RCA can be used with genomic DNA, but the idea of using isothermal amplification is interesting.

I’ll also look into continuous flow microfluidic thermocyclers. Is there anything commercial or is it still mostly R&D?