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Mycoplasmacheck Service

 

 

Easy outsourcing of mycoplasma testing in any cell line

 


Mycoplasmacheck is a qPCR-based service for detection of mycoplasma in cell cultures.

 

 

 

Order Mycoplasmacheck Barcodes:

 

 

 

Why should I test my cell lines for Mycoplasma contamination?

 

As contamination of mycoplasma alters cell-line properties, their unnoticed presence often leads to non-reproducible results. Regular testing with Mycoplasmacheck serves as a rapid, accurate and objective means of checking cells for mycoplasma contamination, ensuring cell lines are kept reliably clean.

Highlights of the Mycoplasmacheck Service:

 

  •  Standardised qPCR test for mycoplasma performed under ISO17025 accreditation
  • Data provided online within 48 hours after sample receipt
  • Test history report for select periods of time can be compiled online at any time

 

 

 

 

 Sample Submission

 

Product Details

Details of our Mycoplasmacheck

 

Full-service package offered by Mycoplasmacheck:

 

  • Detection of all mycoplasma species listed in the European Pharmacopoeia 2.6.7: M. arginini, M. fermentans, M. orale, M. hyorhinis, M. hominis, M. genitalium, M. salivarium, M. synoviae, M. pirum, M. gallisepticum, M. pneumoniae, M. yeatsii, Spiroplasma citri and Acholeplasma laidlawii
  • Identification of more than 100 additional mollicutes
  • Highly sensitive assay capable of detecting 1 international unit (IU) per PCR of the World Health Organization’s (WHO) international standard for mycoplasma DNA (Please see application note)
  • Highly specific test that identifies ten copies of mycoplasma per reaction with no cross-reactivity to E. coli or eukaryotic DNA
  • Use of positive and negative controls to monitor test performance
  • Exclusively licensed from the German Cancer Research Center, (Deutsches Krebsforschungszentrum, DKFZ), Heidelberg, Germany
  • Easy sample drop-off at any Collection Point or ship separately


Technology used for Mycoplasmacheck

 

  • Roche LightCycler 480 II


Deliverables of Mycoplasmacheck

 

  • Data report indicating mycoplasma status, as well as any PCR inhibitors found in the sample
  • The user can compile a PDF document online at any time to view the results from any specific testing period.
  • PDF document serving as an independent certificate of results; suitable for direct submission to journals or granting agencies

 

Sending samples

Sending samples the right way for our TubeSeq Service


Sending samples according to the requirements below helps us to do our job better
and provides you with accurate results.

 


Sample submission

 

  • Use 1.5 ml safe-lock tubes for your templates and primers
  • Do not tape or wrap tubes with parafilm. Safe-lock tubes offer perfect sealing and evaporation protection
  • Label your template with our barcode labels. Either with the prepaid TubeSeq Labels or free Tube Labels
  • Label your enclosed primers with our free Tube Labels
  • Use a water resistant marker for any additional labeling of your tubes

 

 

1

Only use 1.5 ml
safe-lock tubes

 

2

Remove barcode
stricker from film

 

3

Affix barcode sticker
horizontally

 

4

QR-code visible at front so
it can be read by scaner

Sample preparation

Pre-requisites prior testing

  • For maximum sensitivity: Recommendations for cell culture cultivation prior to testing.
  •  Cell lines should be cultured in the absence of mycoplasma-active antibiotics for one week prior to submission. Penicillin and streptomycin do not affect mycoplasma nor do they inhibit MYCOPLASMACHECK.
  • Cell cultures should be maintained in the same media without dilution with fresh media for three days before testing.
  • Cryopreserved cell cultures should be cultured about two weeks prior to sample submission.
  • 80-90% confluence of the tissue culture is recommended for testing. Inhibiting substances may accumulate in >90% confluent cultures.

 

Sample preparation

 

 Sample preparation of cell cultures:

  • Transfer 500 μl of cell culture supernatant from the test cell culture to a 1.5 ml tube. The lid should be sealed tightly to prevent opening during heating.
  • For suspension cell lines, stand culture flasks vertically allowing cells to settle for about 30 minutes prior to removal of 500μL of the supernatant and transfer to a 1.5 ml tube.
  • Boil supernatant at 95 °C for 10 minutes.
  • Briefly centrifuge (5 seconds) the sample at ~13,000 rpm to pellet cellular debris.
  • Transfer 100 – 200 μl of supernatant into a new 1.5 ml tube labelled with a MYCOPLASMACHECK barcode. Do not disturb pellet

 Sample preparation of cell cultures that are prone to PCR inhibition:

 

For adherent cell lines:

 

  • Transfer 500 μl of cell culture supernatant from the cell culture to a 1.5 ml tube.
  • Centrifuge sample for 10 min at >10.000 x g to pellet mycoplasma cells.
  • Discard 450 μL of supernatant without disturbing any pellet.
  • Boil remaining 50μL at 95 °C for 10 minutes. The lid should be sealed tightly to prevent
    opening during heating.
  • Perform DNA extraction using any commercially available DNA extraction kits according to
    the manufacturer ́s instructions.
  • Elute in 30 μL using a 1.5 ml tube labelled with a MYCOPLASMACHECK barcode.

 

For suspension cell lines:

 

  • Centrifuge 500 μL sample for 3 min at >300 x g to pellet eucaryotic cells.
  • Transfer supernatant in new 1.5 ml tube.
  • Continue with 2nd step above.

 

   

 

Testing of other non-cell-culture related material like cell pellets, fetal calf serum or cryo stocks:

 

  • Those materials can be tested after DNA preparation by the customer using commercially available DNA extraction kits. 
  • e.g. QIAamp DNA Mini Kit (QIAGEN) or NucleoSpin® DNA RapidLyse (Macherey-N

Additional Information

Reliably detect all common mycoplasma species

 

Mycoplasma contamination adversely affects 35% of the world’s cell lines.

 

As signs of mycoplasma are easy to miss, only regular monitoring of cell cultures can ensure that research results are valid and reproducible. With its standardised workflows, we can rapidly and reliably detect all common mycoplasma species.

 

 

Table 1. List of mycoplasma species frequently detected in cell cultures. These mycoplasma, all found in the European Pharmacopoeia 2.6.7., are readily detectable with Mycoplasmacheck.


As a qPCR-based assay, the technique is more accurate and sensitive than traditional microbial culture methods and bioluminescence-based methods for mycoplasma detection (Fig1). Besides severely reducing the hands-on time involved in in-house testing, outsourcing the task of mycoplasma detection offers distinct benefits that cannot be matched by do-it-yourself kits (Fig 2).

 

 

Fig. 1. Comparison of three different methods for detection of mycoplasma in cell cultures. The graph demonstrates the superiority of mycoplasma PCR over culture-based or bioluminescence-based (Mycoalert) techniques in analysing cell lines for mycoplasma (adapted from Cytotechnology (2014) 66:861–873)

 

 

Fig. 2. Advantages of using Mycoplasmacheck for mycoplasma PCR detection over in-house PCR kits

 

Order Management

Mycoplasmacheck (MPC) Barcodes

 

  • The format of the MPC Barcodes have changed. They are now 8-digits. Example: MP00000301
  • All "old MPC Barcodes" are still valid and can be used unrestricted
  • The Barcodes can now be found in the "Barcode Management Genotyping" list

Please note: the new Barcodes are already activated. If you still have "older barcodes" (not starting with MP) please activate these prior ordering here

Order updates

 

  • When your samples arrive in our lab an Email will be sent to you and an order will be visible under my orders
  • When samples are finished you will receive a "Partially complete" or a "Complete" Email
    • Partially complete = not all samples are finished
    • Complete = all samples are finished

 

 

 

Literature

 

  

 

 

Related Links

 

 

 

 

 

C. Mario -
07/05/2021 10:06


„Very good quality“


more >>

N. Phillip -
06/05/2021 12:06


„The tube seq service is
competitively priced and
the results arrive within
a reasonable time of 3-5
days...


more >>

A. Petros -
06/05/2021 10:07


„I used to rate the
Sanger Seq with 5 stars,
the late months though
everything got slower and
the sequencing ..."


more >>

S. Dirk -
06/05/2021 12:06


„I am completely happy
with the drop-box overnight
service. Results are
usually available nex day
..."


more >>

 

 

 

C. Mario -
07/05/2021 10:06


„Very good quality“


more >>

N. Phillip -
06/05/2021 12:06


„The tube seq service is
competitively priced and
the results arrive within
a reasonable time of 3-5
days...


more >>

A. Petros -
06/05/2021 10:07


„I used to rate the
Sanger Seq with 5 stars,
the late months though
everything got slower and
the sequencing ..."


more >>

S. Dirk -
06/05/2021 12:06


„I am completely happy
with the drop-box overnight
service. Results are
usually available nex day
..."


more >>

 

 

 

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