MycoRAZOR - Biontex Laboratories

MycoRAZOR ®No Chance for MycoplasmasFor ordering information, MSDS, publications and application notes see www.biontex.comDescription Catalog Number SizeMycoRAZOR ® M040-100 100 mlShipping:–78°CStorage:≤ –15°CStability:Best before: see labelUse:Only for research purposes in vitro, not intended for human or animaldiagnostic, therapeutic or other clinical uses.MycoRAZOR ® is an outstanding effective antibiotic against mycoplasma. It is active at lowconcentrations and against a large variety of mycoplasma. MycoRAZOR ® acts on the proteinsynthesis mechanism by interfering with the ribosome translation of the mycoplasmas as well aswith their transcription.The product is suitable for cell cultures which are infected by mycoplasma and for thedecontamination of primary material.MycoRAZOR ® Manual DE 201312101712 1

Contents1. General Remarks.......................................................................................................................... 31.1 Specifications............................................................................................................................... 31.2 General ....................................................................................................................................... 32. User Protocol ............................................................................................................................... 42.1 Standard Concentration................................................................................................................. 42.2 Further Concentrations.................................................................................................................. 42.3 Application................................................................................................................................... 42.4 Explanatory Notes ........................................................................................................................ 53. Miscellaneous............................................................................................................................... 63.1 Important Information .................................................................................................................. 63.2 Warranty ..................................................................................................................................... 64. Related Products.......................................................................................................................... 72 MycoRAZOR ® Manual DE 201312101712

1. General Remarks1.1 SpecificationsApplicationFormulationAssaysRecommended finalconcentrationRemoves mycoplasmas from contaminated cell culturesAntibiotic mixture in PBSApprox. 300 applications (T75) per 100 ml reagent20 µl/mlpH 4,0 – 4,5OsmolalityEndotoxinSterilityShippingStorage280 – 340 mOsmol/kg< 10 EU/mlTestedOn dry ice≤ -15°C1.2 GeneralThe contamination of cell cultures by mycoplasma remains one of the major problems in biologicalresearch. Most mycoplasma contaminations occur due to the use of animal products like sera ortrypsin. Another possible source of contamination of the cell culture is the laboratory personnelitself.In order to protect you against mycoplasma effectively, a routine examination of the cell cultureand removal of the mycoplasma is necessary. The Biontex products MycoSPY ® (detection) andMycoRAZOR ® (removal) are ideal for these tasks.MycoRAZOR ® Manual DE 201312101712 3

2. User Protocol2.1 Standard ConcentrationThe product MycoRAZOR ® is supplied in a concentration of 0.5 mg/ml in PBS (Dulbecco’s PhosphateBuffered Saline). The amount of MycoRAZOR ® added should always be 1 / 50 of the complete amountof medium in the cell culture flask, giving a working concentration of 10 µg/ml.2.2 Further ConcentrationsIf necessary MycoRAZOR ® can also be applied in higher concentrations:1 / 40 dilution1 / 30 dilution1 / 25 dilution2.3 ApplicationExample for MycoRAZOR ® usage with 1 / 50 dilution:Culture flask T 25 T 75 T 150Culture medium 5 ml medium 15 ml medium 30 ml mediumMycoRAZOR ®100 µl 300 µl 600 µl1. Remove the culture medium from the culture container, wash cells with PBS, and detach cellswith Trypsin or Accutase.2. Count cells; transfer adequate number of cells in appropriate culture vessel with new mediumand an adequate amount of MycoRAZOR ® .3. Cultivate the cells for at least two days in the usual way.4. Repeat step 1 to 3 three times and then check for mycoplasma using MycoSPY ® (or another onPCR based detection system).5. If contamination is still detectable, repeat steps 1 to 3 with increased MycoRAZOR ®concentration (as shown above) and if necessary with longer duration of treatment (increasednumber of passaging steps).Finally check for mycoplasma once using MycoSPY ® .4 MycoRAZOR ® Manual DE 201312101712

2.4 Explanatory Notes1. It is important to check that mycoplasmas have been completely eliminated (e.g. by usingMycoSPY ® ) to prevent the establishment of resistance. As resistance can be built up in thesame way as in all use of antibiotics, complete elimination of mycoplasmas is vital.2. First check whether two passes without the use of MycoRAZOR ® were conducted between thelast use of MycoRAZOR ® and the current test. If this was not the case, dead mycoplasma mayhave been detected, particularly by highly sensitive methods such as PCR. If the minimum timebetween the last application and the test was observed, use MycoRAZOR ® in the next fivepasses of your cells, gradually increasing the dose of MycoRAZOR ® up to a maximum dilution of1/25. Please note that depending on the cell type, toxic effects may result from increasing theconcentration of MycoRAZOR ® . If you observe toxic effects (lower proliferation rate; changes incell morphology) in your cells, apply the last-used dose in the remaining cycles.3. It is essential to test for results of each treatment by using a mycoplasma detection kit!4. The animal products used in cell culture are primary sources of mycoplasma contamination. Toavoid this risk, use only fetal bovine serum (FBS) and trypsin that are guaranteed mycoplasmafree.5. Mycoplasmas belong to the class of Mollicutes and thus lack cell walls; they are resistant tomany antibiotics that attack cell wall synthesis. The user is thus an important source ofcontamination in routine use of this type of antibiotic for cell culture. In this case, non-sterileworking conditions go unnoticed, as the addition of antibiotics prevents the growth of mostbacteria – and thus macroscopic effects – while allowing mycoplasmas to multiply unhindered.6. In addition, cross-contamination from another cell culture is possible. For this reason alwaystest all cultured cells and replace any potentially contaminated cell culture material (medium,FBS, trypsin, buffer, LN2 for storage).MycoRAZOR ® Manual DE 201312101712 5

3. Miscellaneous3.1 Important InformationThis reagent is developed and sold for research purposes and in-vitro use only. It is not intendedfor human therapeutic or diagnostic purposes.3.2 WarrantyBiontex guarantees the performance of this product until the date of expiry printed on the labelwhen used in accordance with the information given in this manual. If you are not satisfied with theperformance of the product please contact Biontex or one of its authorized distributors.6 MycoRAZOR ® Manual DE 201312101712

4. Related ProductsProductMETAFECTENE ®EASY+METAFECTENE ®SI+DOTAPCat.No.T090T100T010ProductTransfectionMETAFECTENE ®PROMETAFECTENE ®FluoRMETAFECTENE ®3D-SCCat.No.ProductCat.No.T040 METAFECTENE ® T021T050 INSECTOGENE T030T110Protein PurificationNi-NTA Agarose R040 Ni-IDA Agarose R010METAFECTENE ®3D-HGHighflow Ni-IDAAgaroseT120R011GST Agarose R030 Co-IDA Agarose R020Highflow Co-IDAAgaroseR021Empty S-Columns P-50S Empty L-Columns P-50L Empty XL-Columns P-50XLEmpty Spin-ColumnsSP-50 Empty Cartridges C010PROTEOfectene ®E010ProteofectionPROTEOfectene ®ABCell CultureE020MycoSPY ® M030 MycoRAZOR ® M040µ-Transfection KitVIK010Microfection Kitsµ-Proteofection KitVIK030µ-Transfection KitVI FluoRK020µ-Proteofection KitVI ABK040Buffer and Positive Controls1x PBS S011 EASY + buffer (10x) EA010SI + buffer (10x) SO010 R-Phycoerythrin S020Visit the Biontex website (http://www.biontex.com) and select your home country to get specific contact and ordering information.MycoRAZOR ® Manual DE 201312101712 7

Biontex Laboratories GmbHAm Klopferspitz 19im IZB82152 Planegg/MartinsriedGermanyPhone: +49 (0)89 3247995-0Fax: +49 (0)89 3247995-2Fax 2: +49 (0)89 32385994E-Mail: contact@biontex.comWeb: www.biontex.com8 MycoRAZOR ® Manual DE 201312101712