For use in quality control/manufacturing process only. Others MycoTOOL Mycoplasma Detection Amplification Kit MycoTOOL Mycoplasma Detection Amplification Kit 3.5.3.19.3.4 05184240001 MycoTOOL, Mycoplasma Detection Amp kit MycoTOOL Mycoplasma Detection Amplification Kit 04038377025773 Reagents, kits 1 kit Not Available CustomBiotech product. Please contact your local representative. Unit of measure is "piece" Mycoplasmas can act as severe contaminants in cell culture. Mycoplasmal cell culture contamination occurs due to contamination from individuals or contaminated cell culture medium ingredients. Mycoplasmas may induce cellular changes, including chromosome aberrations, changes in metabolism and cell growth. Severe mycoplasma infections may destroy a cell line. en Testing of cell culture samples for the absence of mycoplasma. en Sensitive control (10 CFU/mL A. laidlawii): For at least 2/4 replicates a band at 450 bp must be recognized.Negative control of the whole workflow (mycoplasma): For at least 5/6 replicates no band between 420 and 480 bp is recognized.Positive control of the whole workflow (GAPDH): For each sample a band at 150 bp is recognized.Negative control of the PCR (Mycoplasma and GAPDH): For all 4 replicates no band is recognized.Positive control of the PCR (Mycoplasma): For 2/4 replicates a band is recognized at 360 bp. en The MycoTOOL Mycoplasma Detection Amplification Kit is an in vitro nucleic acid amplification test optimized for the detection of mycoplasmas in CHO cell culture according to the Guideline for Mycoplasma NAT described in Chapter 2.6.7 of the European Pharmacopoeia with respect to specificity, sensitivity/detection limit, and robustness.The kit can also be used for cell cultures derived from several other cell lines. The performance criteria need to be established by the user.As internal control of the lysis and sample preparation efficiency, the kit includes a primer pair for the GAPDH housekeeping gene of CHO cell DNA to be used with a cell culture sample 5 × 106 cells/mL. For cell culture samples with 5 × 106 cells/mL to 1 × 108 cells/mL CHO, use a housekeeping gene for SP2 and CHO.Prior to PCR amplification, the risk of amplicon contamination is reduced by using Uracil-N-Glycosylase enzyme. It recognizes and catalyzes the destruction of DNA strands containing deoxyuridine (amplicon), but not DNA containing deoxythymidine (sample).In combination with this kit, a manual sample preparation with the QC Sample Preparation Kit is recommended. For more information related to its content, refer to the corresponding Instructions for Use.A sample preparation protocol with the QC Sample Preparation Kit for cell culture sample with a density of <5 × 106 cells/mL and also a protocol for high cell density (5 × 106 cells/mL to 1 × 108 cells/mL, derived from SP2 and CHO) is described in section, Protocols. en