No More Mycoplasma

October 06, 2016

University of Kansas Medical Center
Pathology
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Sigma Aldrich

Product Name:

Lookout Mycoplasma Elimination Kit

Catalog Number:

MP-0030

The specific goal was to eliminate mycoplasma from my cell cultures, which I think is clear. I chose this product because I received an email discussing mycoplamsa elimination options from Biocompare. Also, the kits were complete and appeared to be user-friendly.

Experimental Design and Results Summary

Application

Cell Culture

Starting Material

Contaminated cultures, grown in 100 mm dishes in RPMI supplemented with 10% FBS.

Protocol Overview

Passage cells at low density (single cells) in media containing Step I elimination antibiotic. Allow cells to return to 80% confluence, passage as usual with Step II elimination antibiotic (3 rounds of treatment). Re-test for mycoplasma before resuming assays.

Tips

Avoid adding other antibiotics (e.g. pen/strep) to cultures as this may be toxic to cells. Sigma recommends reducing FBS concentration in media during treatment.

Results Summary

Following a total of 4 treatments (Step I - once, Step II - 3 times), my cell cultures were mycoplasma free as tested both using the Sigma detection kit and by sending out for third party testing.

Additional Notes

N/A

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Summary

The Good

Quick, easy!

The Bad

A little expensive, especially for multiple cell lines. Also, its unclear if the antibiotic treatment affects cells

The Bottom Line

This is a good mycoplasma elimination tool. I would recommend, especially for hardy cells. There may be issues with use in cell lines that are generally more sensitive.

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