Hi,

I used Light Cycler 480 Probes Master Mix with EvaGreen dye in Real Time PCR protocols, and I got satisfactory results. However, I am not sure this mix is suitable for conventional PCR, because it is designed for Real Time PCR.

I generally use SsoFast EvaGreen Mastermix (Biorad) or for very pure DNA samples Bioline No-Rox Mastermix. The Biorad MM also works with "quick and dirty" approaches, e.g. cell lysates; but it is more expensive. 

Both companies ususally grant special offers for academics. 

Thanks, I have had contact with Biorad about SsoFast EvaGreen Mastermix. They didn't recommend this mix for gene expression. Bioline No-Rox Mastermix is with SYBR. I prefer a mix with EvaGreen.

Hi

It is not EvaGreen but we've used Resolight from Roche, seems to be OK. Gave good first derivative melting curves for genotyping. We've found that SYTO9 is better than EvaGreen for most of our assays (not sure of the price comparison though). We've been using the Roche faststart taqman mastermix with SYTO9 since we can also do taqman and only need to buy the 1 mastermix. For assays that don't like the high Mg we make our own mastermix with GoTaq or Amplitaq Gold. At the end of the day you need to use a mastermix that works for your assay, if you are using an intercalator then it has to have minimal non-specific amplification if you want to do quantification

We have chosen for 5x HOT FIREPol EvaGreen qPCR Mix Plus (without rox) from Solis BioDyne, and meanwhile good experience with it.

Judith Hendriks

I tried it with the same conditions in the protocol but the result was not good.... do you recommend any modification in the protocol?