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MycoTOOL PCR Mycoplasma Detection Kit

For use in quality control / manufacturing process only.

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Order information
MycoTOOL PCR Mycoplasma Detection Kit material number and pack size:
Material Number Pack Size
05184240001 1 kit: MycoTOOL Mycoplasma Detection Amplification Kit
05184592001 1 kit: MycoTOOL Mycoplasma Detection Prep Kit
Unit of measure is "piece".
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Overview
For the testing of cell culture samples for the absence of mycoplasma.
The MycoTOOL Mycoplasma Detection Amplification Kit supplies all reagents for the amplifications step.
Mycoplasma can act as severe contaminant in cell culture. Mycoplasma 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.
  • Save time. Replace traditional 28 days culture testing with the rapid MycoTOOL PCR Test.
  • Speed up your validation. Rely on published Roche validation data.
  • Detect traces of Mycoplasmas. The sensitivity is 1 CFU/mL.
  • Minimize the risk of false negatives and false positives. The matrix lysis control eliminates risk of undetected intracellular Mycoplasma. The diluted GAPDH and positive control identify potential PCR inhibition. False positives are prevented by nucleic acid free reagents, tested according to Roche's current ultrasensitive Quality Control procedures. Uracil-DNA Glycosylase pretreatment is performed to prevent contamination by PCR carryover.
  • Detect the broad panel of Mollicute species. A universal primer design covers a whole range of Mycoplasmas.
  • Achieve process security. Rely on test results using a consistent kit quality and established change control procedures."
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Application
Highly sensitve PCR test (Block PCR analysis via gel) for the detection of Mycoplasmas, accepted for validation according to the E.P.2.6.7 directive. this test is suitable  for release testing and in-process control. it can replace culture and indicator cell tests. Use the MycoTOOL test in the field of biopharmaceutical and vaccine production, cell therapy and transplantation.
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Specification
Using CHO cells as reference cells for spiking experiments we recommend the CHO-K1 (ATCC CCL-61) cell line. The cells should be grown in DMEM/F12 (containing 10% FCS) : RPMI (2:1) according to ATCC recommendation. After trypsination cells should be diluted to a concentration of 5 ×106 cells/mL using the original growth medium.
Sensitive control (10 CFU/mL A. laidlawii): for at least 2/4 replicates a band at 450 bp must be recognized: 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 (GAPDH): for each sample a band at 150 bp is recognized: Corresponds specification
Negative control of the PCR (Mycoplasma and GAPDH): 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 to -25°C within specification range for 24 months.