The cell culture supernatant (sample) can be stored for several days at 4 °C before using the kit to determine if mycoplasma is present. For long-term storage, the samples can be stored at -20 °C, either in their native state or after heat inactivation.
MP0025
Venor™ GeM Mycoplasma Detection Kit, PCR-based
Synonym(e):
PCR mycoplasma detection kit
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NACRES:
NA.75
UNSPSC Code:
12352207
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Unterstützung erhaltenusage
kit sufficient for 25 tests
packaging
pkg of 1 kit
storage condition
dry at room temperature
technique(s)
PCR: suitable
application(s)
detection
microbiology
storage temp.
2-8°C
Quality Level
Application
General description
The Venor™ GeM Mycoplasma Detection Kit utilizes the polymerase chain reaction (PCR), which was established as the method of choice for highest sensitivity in the detection of Mycoplasma and Acholeplasma contamination in cell cultures and other cell culture derived biologicals. Detection requires as little as 1–5fg of mycoplasma DNA corresponding to 2–5 mycoplasma per sample volume. The primer set is specific to the highly conserved rRNA operon, or more specifically, the 16S rRNA coding region in the mycoplasma genome. This allows for detection of all Mycoplasma, Acholeplasma and Ureaplasma species tested so far, which are usually encountered as contaminants in cell cultures. Eukaryotic and bacterial DNA are not amplified by this kit.
Other Notes
Does not include Taq Polymerase. Optimized for use with D9307, Taq DNA Polymerase
Preparation Note
The kit components are stable during shipping at ambient temperature. Upon receipt, store at 2–8 °C. After rehydration of the primer/nucleotide mix, the positive control, and the internal control, store below –20 °C and avoid repeated freezing and thawing. For repeated testing of low sample numbers, primer/nucleotide mix and controls should be aliquoted after rehydration. By following these recommendations, the kit is stable until the expiration date stated on the label.
Legal Information
Venor is a trademark of Minerva Biolabs GmbH
Nur Kit-Komponenten
Produkt-Nr.
Beschreibung
- Positive Control
- Negative Control
- PCR 10X Reaction Buffer 500 mL/vial
- Primer/Nucleotide Mix 1 mL/vial
Lagerklasse
10 - Combustible liquids
wgk
WGK 3
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Verwandter Inhalt
Instructions
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Can the cell culture supernatant be stored for a period of time before using the kit to determine if mycoplasma is present?
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Can I freeze the samples to collect several before using them?
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Samples should be cell supernatants when possible. Cell pellets are not recommended. The high concentration of DNA in the pellet is often responsible for the non-specific bands. If cell pellets are the only material available for testing, then centrifugation is necessary following sample boiling in order to remove all cell debris.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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The sample required for VenorGeM® Mycoplasma Kit, Product MP0025, is a cell culture supernatant. Is this from adherent or suspension cells?
1 answer-
The mycoplasma assay kit can be run on supernatants from either adherent or suspension cells. The centrifugation step during sample preparation will remove any cellular debris to limit PCR inhibitors.
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When using Product MP0025, VenorGeM® Mycoplasma Kit, there is inhibition in the sample (no internal control band). Can the sample be diluted and the assay repeated?
1 answer-
Sigma Aldrich does not recommend diluting the sample and re-assay as this can result in false negatives (sample diluted below the detection limits of the assay). Sigma Aldrich recommends that PCR inhibitors be removed using a DNA extraction with commercially available kits. (Product Codes G1N10, G1N70, NA2000).
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Product MP0025, VenorGeM® Mycoplasma Kit, does not contain TAQ. What DNA polymerase should be used?
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Any DNA polymerase with a 3’-5’ exonuclease activity is NOT recommended. These include Tli, Pfu,and Pwo. Those DNA polymerases that can be used include Taq, Tfl, and Tth. The kit has been optimized for use with Product No. D9307.
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What is the difference between Product No. MP0025 (VenorGem Mycoplasma Detection Kit) and Product No. MP0035 (Lookout Mycoplasma Detection Kit)?
1 answer-
The assay kits have a similar range of species detected (see product insert for listing). For Product No. MP0025, the components are supplied as individual reagents. For Product No. MP0035, the components are supplied in reaction tubes, already mixed and ready for addition of sample and Taq. The internal control for Product No. MP0035 is at 481 bp rather than 191 bp as found in Product No. MP0025.
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Can a cell lysate be used in the VenorGeM® Mycoplasma Kit, Product MP0025, assay?
1 answer-
A cell lysate can contain PCR inhibitors. Sigma Aldrich recommends that PCR inhibitors be removed using a DNA extraction with commercially available kits. (Product Codes G1N10, G1N70, NA2000).
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