In my experience an internal control is needed as numerous inhibitors are present in stool samples. I have used mCMV as an internal control.

not real time, just RT-PCR..

thank you Marc!

Hi Renata,

In your case internal control would be necessary mainly to know that your PCR worked, and you didn’t get a false negative result.

Some detection PCR kits arrive with internal controls for that purpose like mycoplasma detection kits.

Dear Renata

Internal controls were used for qPCR. In RT-PCR you must run two control:

1. Positive control (that contain known RNA segment that you looking for)

2. Negative control (That have all reagent except cDNA)

With help of these two controls, you can check the PCR working and reagent purity.

Also you must use ladder to identify the band weight.

Cheers

Hello,

As previously said, controls are mandatory if you want to insure your result, especially in a complex matrix like stool. Inhibitors are present in your sample and you have to check whether there is an influence on your results, so use a positive control in your PCR and if inhibited partially or totally, that means that your sample comprises inhibitors and have to be diluted if you want to have a better signal...

Cheers