Hi,

I was wondering if anyone has a master mix protocol that acts as a substitute for the MicroSEQ 500 16S rDNA PCR kit from Thermo (https://www.thermofisher.com/order/catalog/product/4348229) . We do microbial ID's in our lab and I figured that there's some protocol out there to make the master mix instead of spending ~$750 per kit. I figure that it has universal primers (16S F & R 500bp product), but I'm not sure on buffer/buffer concentration (if using taq, GoTaq 10x?), dNTP concentration, MgCl concentration, and the polymerase used (taq?). I've tried googling it and I can't really find anything. Everything is just using the kit itself. I'd be okay with using the Prepman Ultra kit, but I'd like to make our own master mix. Any help would be greatly appreciated.

Thanks!