Cambio - Excellence in Molecular Biology

Cell Culture Contamination Control

Cell Culture Contamination Control: Detection

Mycoplasma Detection

As an efficient contamination control measure we recommend frequent routine testing of cell culture materials with Venor®Gem Mycoplasma Detection Kits. These PCR- and qPCR-based mycoplasma assays allow fast (within a few hours), simple, effective and sensitive detection of mycoplasma-contaminated cell cultures in any standard molecular biology laboratory.

Regular mycoplasma testing and control procedures can significantly help in avoiding the costly consequences of contamination ensuring safe biopharmaceuticals and reliable scientific results.  Many scientific journals currently do not accept submissions of cell-based studies without evidence that mycoplasma testing has been performed.

To meet different customer needs Minerva Biolabs has developed a range of Venor®GeM mycoplasma detection assays using conventional PCR or qPCR methods which enable rapid and reliable identification of mycoplasma in cell cultures, culture supernatants and tissue culture media.

Conventional PCR

  • Venor®GeM Classic allows fast, reliable and time-saving routine monitoring of mycoplasma contamination in cell culture supernatants, media and biopharmaceuticals in research and industry by conventional PCR.

  • Venor®GeM OneStep is a kit for direct detection. It requires minimal pipetting, the kit includes all reagents required for PCR in a ready-to-use lyophilized reaction mix. Only samples or controls need to be added prior to PCR.

  • Venor®GeM Advance is designed for direct detection. Designed to reduce the total assay time and the pipetting steps, the kit contains PCR strips of tubes, each pre-dispensed with all reagents necessary for each PCR reaction, including the polymerase. For additional convenience, the gel loading buffer and dye are already included in the reaction buffer. After PCR, the products can be loaded directly on an agarose gel.

Real-time PCR (qPCR)

Our qPCR kits are TaqMan®-based qPCR Assays with labelled probes:  FAM® channel (mycoplasma) and HEX® channel (internal control to check for PCR inhibition from contaminants in sample).  So not only does the qPCR approach remove a lot of hassle, but the internal controls also give you reassurance that your negatives are not false.

  • Venor®GeM qEP utilises quantitative, real-time PCR (qPCR) for fast (approx. 3 h) and reliable screening of cell culture supernatants for mycoplasma contamination. The kit can be used in combination with any type of real-time PCR cycler able to detect the fluorescence dyes FAM® and HEX®. For clinical diagnostics in which human samples are tested we recommend using this kit as it conforms to EP 2.6.7, USP <71> and JP G3. It is suitable for both research and industry where certified compliance is needed or desirable. 

  • Venor®GeM qOneStep is a kit based on a real-time PCR (qPCR) assay for rapid, robust and sensitive detection of mycoplasma contamination. The detection procedure can be performed within 3 hours. Primers, nucleotides, probes, polymerase and an internal amplification control are provided as a ready-to-use, lyophilized reaction mix. This kit is a very straightforward and is highly suitable for academic labs and industrial applications where certified compliance of kits is not required.


Venor®GeM-qEP Mycoplasma Detection Kit

Venor®GeM qEP utilizes quantitative, real-time PCR (qPCR). The kit can be performed with any type of real-time PCR cycler able to detect the fluorescence dyes FAM™ and HEX™. The protocol provided is preferred for fast and reliable screening of cell culture supernatants most applicable in research and development. The detection procedure can be performed within 3 hours. Important note: Please see the note section for important information on the use of the supplied positive control

Minerva Biolabs

Catalogue No.DescriptionPack SizePriceQty
11-9025Venor®GeM qEP25 rxns £329.00 Quantity Add to Order
11-9100Venor®GeM qEP100 rxns £1,086.00 Quantity Add to Order
11-9250Venor®GeM qEP250 rxns £2,205.00 Quantity Add to Order
11-9905Internal Control DNA extra for VenorGem qEP4 vials £22.00 Quantity Add to Order

Description

Venor®GeM qEP

Mycoplasmas are specifically detected by amplifying a highly conserved rRNA operon, or more specifically, the 16S rRNA coding region in the mycoplasma genome. The mycoplasma-specific amplification is detected at 520 nm (FAM™ channel).The kit includes primer and FAM™ labeled probes which allow the specific detection of all Mollicutes species so far described as contaminants of cell cultures and media components. Eukaryotic DNA is not amplified by this primer/probe system.
False negative results due to PCR inhibitors or improper DNA extraction are detected by the internal amplification control which can be added to the PCR master mix. The amplification of the internal amplification control is detected at 610 nm (HEX™ channel).

Important note:

Please see the note section for important information on the use of the supplied positive control

Type of PCR

TaqMan® based qPCR Assay with FAM™ and HEX™ labeled probes

Recommended Use / Scope

Venor®GeM qEP is used for direct detection of Mollicutes (Mycoplasma, Acholeplasma, Spiroplasma) contamination in cell cultures and cell media components.

Kit Components

lyophilized primer/nucleotide/probe/polymerase/mix in aliquots of 25 tests
rehydration bufferlyophilized internal amplification control
lyophilized positive control
PCR grade water

Package Sizes

  • Cat. No. 11-9025 25 tests
  • Cat. No. 11-9100 100 tests
  • Cat. No. 11-9250 250 tests

Result Evaluation

cycler based, real-time PCR

Required Consumables

PCR reaction tubes
Optional for process validation and EP 2.6.7 compliant testing:
Internal Control DNA extra (4 vials for 300 µl each of internal amplification control; Cat. No. 11-9905)
10CFU™ Sensitivity Standards available for all EP 2.6.7 listed mycoplasma species

Required Lab Devices

qPCR cycler with FAM™ and HEX™ filter
variable microliter pipettes
benchtop centrifuge for 1.5 ml reaction tubes

Shelf Life and Storage

Components can be stored at 2 to 8 °C for at least 12 months. After rehydration the reagents must be stored at -18 °C.

EP 2.6.7 Compliance

Yes.Please note that validation data are provided for information purpose only. EP 2.6.7 clearly states “Where commercial kits are used …, documented validation points already covered by the kit manufacturer can replace validation by the user. Nevertheless, the performance of the kit with respect to its intended use has to be demonstrated by the user (e.g. detection limit, robustness, cross-detection of other classes of bacteria.”. Please feel free to contact us if you need further assistance.

Fig. Amplified dilution series of Mycoplasma fermentans, performed on a
Corbett RotorGene®6000.

 

 

?

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Protocols

Venor®GeM qEP protocol

Quick Visual Protocol

Please note: all protocols off site are the responsibility of the products supplier

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

References

For a list of citations please click here

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Notes

Important notes on the use postive controls in the VenorGem™ QEP kit

The Positive Control comes in one vial. After rehydration, this reagent is stable for frequent thawing/freezing cycles. However we strongly recommend that customers use  DNase-free pipette tips and a clean surrounding. One bacteria particle contaminating the reagent is able to digest all the DNA during the thawing phase.

Aliquoting the rehydrated Positive Control can help avoid this as long as the following criterion are met:
1. Make aliquots in PCR tubes (they are usually clean, DNase-free, low-binding tubes, low surface area).
2. Never ever use reaction tubes or other larger vials, as they absorb the minute amounts of DNA
3. keep the thawing time short. Freeze asap.

Required consumables:

PCR reaction tubes

Optional for process validation and EP 2.6.7 compliant testing:

Internal Control DNA extra (4 vials for 300 µl each of internal amplification control; Cat. No. 11-9905)

10CFU™ Sensitivity Standards available for all EP listed mycoplasma species

 

Required lab devices:

qPCR cycler with FAM™ and ROX™ filter

variable microliter pipettes

benchtop centrifuge for 1.5 ml reaction tubes

 

Shelf life and storage:

Components can be stored at 2 to 8 °C for at least 12 months. After rehydratisation the reagents must be stored at -18 °C.

 

EP 2.6.7 Compliance:

Yes.

Please note that validation data are provided for information purpose only. EP 2.6.7 clearly states “Where commercial kits are used …, documented validation points already covered by the kit manufacturer can replace validation by the user. Nevertheless, the performance of the kit with respect to its intended use has to be demonstrated by the user (e.g. detection limit, robustness, cross-detection of other classes of bacteria.”. Please feel free to contact us if you need further assistance.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications & Benefits

 

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200