Mistake during mycoplasma-pcr - (Dec/09/2008 )

Hi everybody,

I've got a stupid question:
Afew month ago we did mycoplasma pcr, but forgot to heat the samples before the pcr.
What is this boiling for? Should the heating during the pcr compensate that?

Thanks a lot

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AFAIK heating prior to PCR is to denature the DNA. Most PCR programs have a step in them (5-10 min 95 C or so) to do this. We never heat the samples prior to (myco) PCR, but have such a step in the PCR program.

If your PCR program has such a step I would not worry too much about it. Furthermore, you could check the positive and negative controls to see if the PCR worked.

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Great. I have been hopeing for something like that. Thank you.

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Dear gw3,

My favourite subject yet again appears......oh dear !!!!!!!!!

Your first mistake is using PCR as a method for mycoplasma detection.........my reasons for this statement is backed up by the following arguements:

PCR IS NOT FDA APPROVED AS A METHOD FOR MYCOPLASMA DETECTION......this statement means that any drug or vaccine you might take IS MYCOPLASMA TESTED......BUT NOT BY THE PCR METHOD.


ATCC....I think the worlds largest commercial supplier of cell lines, mycoplasma test all their cell lines. You know what is coming next: THEY NO LONGER DO OR OFFER PCR AS A METHOD FOR MYCOPLASMA DETECTION.


I have had groups in the Institute I work in stating that their cells are "mycoplasma free" after testing them using PCR. They do not listen to Uncle Rhombus. 6 months down the line send their cells of for proper testing, only to find that they are massively contaminated with mycoplasma. The PCR test regularly gives many FALSE NEGATIVES.

3 statements of fact which you should note with EXTREME CAUTION.


What is the best tests and that are FDA approved.......Search under Rhombus and find out

Regards

Rhombus

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