Eliminate Free DNA, Only Detect Viable Organisms
The Grim Reefer Free DNA Removal Kit eliminates free DNA from samples prior to qPCR analysis. This provides a quick, accurate, and affordable solution to a problem that has haunted labs using DNA-based methods. Use Grim Reefer to ensure DNA from dead organisms does not inflate quantitative results from PathoSEEK® Microbial Safety Testing Assays.
Live-Dead: A Problem that Haunts qPCR Testing Labs
One of the common objections to using qPCR for microbial testing is the fact that the method does not distinguish between live and dead DNA. qPCR primers and probes will amplify any DNA in the sample that matches the target sequence, regardless of viability. In the past, labs have had to use costly, time-consuming, and possibly hazardous methods to solve this live-dead problem.
Our proprietary Grim Reefer Free DNA Removal method is safer, faster, and better than other live-dead solutions.
The Grim Reefer Process
Add the Grim Reefer buffer and enzyme to the sample and incubate at 37C for 10 minutes to degrade any dead of free DNA. After incubation, add MGC lysis buffer to the sample and mix well. The lysis buffer inactivates the Grim Reefer Enzyme while also lysing the viable cells. After the Lysis step, add the Grim Reefer Positive Control, which contains a known amount of free DNA, to ensure the enzyme is deactivated. If the Grim Reefer Positive Control appears in the qPCR results, you know the Grim Reefer Enzyme was properly deactivated.
Add Grim Reefer Enzyme and Buffer to homogenized sample and incubate for 10 minutes.
Grim Reefer Enzyme eats any free DNA present in the sample
Add MGC Lysis Buffer
Lysis Buffer deactivates Grim Reefer Enzyme and lyses viable cells to expose DNA
Add Grim Reefer Positive Control
Positive Control is Free DNA used to show the Grim Reefer Enzyme has been deactivated