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.