Hello
I have been asked to test mycoplasma contanimation in some cell culture samples by the adjacent lab using PCR. The lab has been getting their cultures tested outside and know they are contaminated, however they want to be able to do it in the building and asked me to set up the PCR and test their samples.
I am worried about my PCR space and pipettes and other equipment. I work with precious human DNA and RNA samples and don't want this mycoplasma testing to contaminate everything else too.
Can the mycoplasma contamination extend to PCR bench spaces/pipettes and other equipment in use? esp the thermocycler? Or am I stressing over nothing?
Please advise.
What's your point exactly?
How are you working with other samples and are you sure that you don't have contamination?
There is some tips that you have to follow. The simplest one is using neg and pos control in each PCR run.
Regards.
DEar Shruti
mycoplasma is a bacteria and therefore if it is true that its contamination could be easly spreaded beetween cultures in the same incubator it can be killed by cleaning well all the surfaces with isopropanol 30%, etoh70% or sodium ipocloride as well as the most part of other bacteria therefore i think you stress little bit to nuch youself. It is not a phage that it is a DNA the therefore need storngest traeatment to be removed
You can find a possibile protocool for mycoplasma test in my blog: PRoteoCool ( https://proteocool.blogspot.com/p/page-5-microbiology.html )
in our case to minimize the risk of mycoplasma speading, we perform the first part of sample preparation inside a laminar flow chamber that could be later decontaminated by UV. There we collect a sample from the culture into an eppendoft , it is closed, trasfered outside and therefore we perform heat denaturartion. After heat denaturation theoretically the bacteria are died and theferore you can prepare your PCR with out many concerns.
Of course if you are worried about contamination at the end you can clean all the sufrace well as already discussed previoosly, use tips with filter and clean also the pipettes or sterilize it by autoclaving, but really i think that it is not so important.
good luck
Manuele
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