|The Venor®GeM Mycoplasma PCR Detection Kits allow rapid
and reliable detection of mycoplasma in various in situ
biologicals including cell cultures and virus stocks.
Tips and hints to optimise your mycoplasma testing:
- Internal control – this is a separate amplification that takes place within a test PCR reaction. It is an outstanding validation tool showing that the PCR is amplifying product as expected and can help rule out false negatives. In the case of the VenorGeM kits the internal control is designed to compete weakly with the target for the amplification. Consequently if your target is strongly amplified, you don’t need to worry if the internal control fails to amplify in this individual reaction! However, if samples fail to show both amplifications (target and internal control), you have reason to believe that your PCR reaction did not work properly in your experimental conditions and that that sample might be a false negative.
- Positive control – another important control to validate your PCR. Large amounts of amplifiable template DNA will ensure further reliable verification of PCR performance. One thing you might want to consider is to handle your positive control DNA in a separate area with separate pipettes to avoid contamination of your PCR testing area with considerable quantities of amplifiable template DNA.
- Handling positive control DNA – Positive control DNA is designed to be at the right concentration so that amplification is not excessive and this allows the presence of inefficient amplification to be picked up by the user. So when you rehydrate your positive control in the buffer supplied aliquot it immediately into low-binding plastic tubes, prepare small aliquots and freeze immediately – and yes, avoid freeze thawing.
Negative control – in certain cases PCR grade water is a sensible choice as negative control. However in the case of VenorGeM kits you might alternatively want to control your sample matrix, using the very same solution/diluent present in your samples. Using a sample of the medium that you are using allows you both to check it for mycoplasma contamination, and for potential interference of its components with the PCR reaction, ruling out once again false negatives.
For more information, please visit us at www.cambio.co.uk or call us on 01954 210200