Hi, I am looking to buy a kit for earliest detection of bacteria and fungus in cell culture (mESCs/ MEFs). I know these contaminants are easily detected by eye/ under the microscope. But I've been having recurrect issues of contamination since a year, and wondered if a kit could help me detect them much earlier than a week or two in culture.
Many thanks,
Pooja.
For mycoplasma you can use some DAPI stain. Just take an aliquot of your TC flasks at regular intervals and add a drop of DAPI. Then observe under the microscope. If you have tiny little bright almost star-like objects that would indicate mycoplasma.
Alternative you can opt for a kit.
However, for re-current contaminations I would really make sure the working solutions and media are filtered before use, incubators and hoods are cleaned regularly (we clean ours weekly). Have dedicated labcoat for your TC work. Use sterile plasticware etc
good luck
There are kits that can be commercialy procured from many compnies for instance Lonza Has a kit to detect mycoplasma using colorimetric analysis where as some labs do that using PCR
HI
Below link provides the info you asked.
https://www.thermofisher.com/order/catalog/product/C7028
hope this helps
For mycoplasma you can use some DAPI stain. Just take an aliquot of your TC flasks at regular intervals and add a drop of DAPI. Then observe under the microscope. If you have tiny little bright almost star-like objects that would indicate mycoplasma.
Alternative you can opt for a kit.
However, for re-current contaminations I would really make sure the working solutions and media are filtered before use, incubators and hoods are cleaned regularly (we clean ours weekly). Have dedicated labcoat for your TC work. Use sterile plasticware etc
good luck
Hello
Divide your cells into three tubes: 0.5 mL for bacterial testing, 2 ml for fungal and 2.5 ml for Mycoplasma testing
For bacterial testing: Add the 0.5 ml to 4.5 ml of Tryptose Phosphate Broth. Incubate at 37°C for 48 hours. Check for growth
For fungal testing: Inoculate a Sabouraud plate with the 2 ml of culture. Culture for 14 days at room temperature. Check for growth
For Mycoplasma testing: SPIN down the 2.5 ml of culture and aspirate until 200 ul of media remains. Extract DNA using a DNeasy kit following the manufacturer’s instructions. Test for the presence of Mycoplasma using a PCR-based detection method
Find links here for more details , Good Luck
http://www.ncbi.nlm.nih.gov/pubmed/8070453
http://www.bio-link.org/home/sites/files/instructor_manualv8.pdf
http://www.invivogen.com/docs/Insight200511.pdf
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4112089
Hi Pooja,
For mycoplasma detection there a easy, comlete PCR kit from Biological Industries http://www.bioind.com/mycoplasma-detection-kit
You can also treat precious cultures with antibiotics against mycoplasma, if the PCR test comes out positive. http://www.bioind.com/page_14202
Hope this helps
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