MycoTOOL PCR Mycoplasma Detection Kit for High Cell Density
For use in quality control / manufacturing process only.
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MycoTOOL PCR Mycoplasma Detection Kit for High Cell Density material number and pack size: Material Number Pack Size 06537456001 1 kit (consists of 2 subkits)
For the testing of cell culture samples for the absence of mycoplasma.
Mycoplasma can act as severe contaminant in cell culture. Mycoplasmal cell culture contamination occurs due to contamination from individuals or contaminated cell culture medium ingredients. Mycoplasma may induce cellular changes, including chromosome aberrations, changes in metabolism and cell growth. Severe mycoplasma infections may destroy a cell line.
The MycoTOOL PCR Mycoplasma Detection Kit is an in vitro nucleic acid amplification test optimized for the detection of mycoplasma in CHO cell culture according to the Guideline for Mycoplasma NAT described in chapter 2.6.7 of the European Pharmacopeia with respect to specificity, sensitivity/detection limit and robustness.
- The modified MycoTOOL PCR Mycoplasma Detection Kit for High Cell Density is suitable for processing samples with high cell density in the range of 5x106 tc/mL to 1x108 tc/mL, derived from SP2 and CHO.
As internal control for the lysis and sample preparation efficiency the additionally shipped primer pair Housekeeping Gene Primer Mix, SP2/0, CHO matching a housekeeping gene of SP2 and CHO is recommended to be used.
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 samples containing deoxyuridine (amplicon), but not DNA containing deoxythymidine (sample).
Sensitive control (5 CFU/mL A. laidlawii): For at least 3/6 replicates a band at 450 bp must be recognize: Corresponds specification
Negative control of the whole workflow (mycoplasma): For at least 5/6 replicates no band between 420 and 480 bp is recognized: Corresponds specification
Positive control of the whole workflow (ß Aktin): For each sample a band at 166 bp is recognized: Corresponds specification
Negative control of the PCR (Mycoplasma and ß Aktin): for all 4 replicates no band is recognized: Corresponds specification
Positive control of the PCR (Mycoplasma) for 2/4 replicates a band is recognized at 360 bp: Corresponds specification
Stability: At +15°C to +25°C within specification range for 18 months.