One-Step Reverse Transcriptase PCR Kits...but which one?

By custom and practice my lab has been using the One-Step Reverse Transcriptase-PCR kit from Invitrogen (aka Life Technologies, aka Thermo Fisher scientific). It works just fine, but in these austere times (I’ve heard even Post-Docs are having to share their ideas) and being under pressure to keep to my departmental budget, I’ve looked at some alternatives as the Invitrogen kit is relatively expensive.

Thermo do a Verso kit which is cheaper and affymetrix have a well-priced kit. The thing that puts me off the Thermo kit is the accompanying blurb…basically it’s a ‘put the things in the tube and do the the PCR bonehead’ …kind of guide. On the other hand, the guide from Affymetrix strokes your hair and hums to you whilst you’re doing the prep.

Aside from the merits of the one step/two-step protocols, has anyone got some other suggestions for a kits that they find work (always a bonus) and that are well priced?

Although we are based in Germany, we have access to most companies here…hey we’ve even had Walmart here for a while, but people thought there was something suspicious about being greeted at the door!

Any suggestions would be most welcome…

Steve

Although maybe not the most direct answer, we usually use two-step RT kits, either Superscript III (Invitrogen) or the iScript kit (BioRad). The latter is RNase H+ which can be an advantage for qPCR as it will remove mRNA following amplification, but might make cloning out a cDNA more challenging. I usually quarter the recommended reaction to save $, if trying to clone a gene, and use either KOD polymerase or try homemade Pfu. Not sure if that helps, or if you strongly prefer the one-step kits :slight_smile: