PCR test kit for rapid, highly sensitive detection of mycoplasmas in cell cultures
Catalogue Numbers:
- M020-025 (25 Assays)
- M020-050 (50 Assays)
Sizes:
- 25 Assays (M020-025)
- 50 Assays (M020-050)
HIGHLIGHTS:
- Detects all cell-culture-relevant mycoplasma strains
- Highly sensitive ( >80 copies of the mycoplasma genome)
- Master Mix with loading buffer and tracking dyes allows for easiest protocol (see workflow)
- Fast results in less than 3 hours
Product Specification
| Application | PCR Kit for detection of mycoplasma contamination in cell culture |
|---|---|
| Content | Lyophilized Master Mix with integrated loading buffer and tracking dyes, internal control and PCR-grade water |
| Assays | 25 or 50 (2 x 25) assays per kit |
| Shipping | Ambient temperature |
| Storage | ≤ –15°C |
| Shelf life | 1 year (from date of delivery) |
| Product Group | Mycoplasma Free Cell Culture |
| Sensitivity | > 80 copies of the mycoplasma genome |
A number of options are available for conclusive detection of the presence of mycoplasmas in contaminated cell cultures. A commonly but unreliable method is the DAPI staining of cells followed by microscopic analysis. However, PCR based methods give the most reliable results for routine checks. Biontex (AFSBio's supplier) MycoSPY® and MycoSPY® Master Mix detection kits are rapid, reliable and highly sensitive methods of PCR-based mycoplasma detection in cell cultures.
The MycoSPY® Master Mix contains a complete range of components for PCR analysis. Each mycoplasma test kit additionally contains an internal control and PCR-grade water for rehydrating the Master Mix.
Workflow
The Master Mix with integrated loading buffer and tracking dyes is designed for maximum ease of pipetting and direct loading of the agarose gel with the PCR samples:
More than a quarter of all animal cell cultures are contaminated with mycoplasmas / mollicutes. The strains most commonly found in cell cultures, with a total probability of 94%, are: M. fermentans (47%), M. hyorhinis (19%), M. orale (10%), M. arginini (9%), A. laidlawii (6%) and M. hominis (3%). In addition, the following strains were found with lower probability: M. gallisepticum, M. pneumoniae, M. salivarium, M. synoviae and S. citri. All these Mollicute strains, in addition to a variety of other strains, are detected by MycoSPY® Master Mix. BLAST analysis was used to verify of the primer specificity. The list shows only Mollicute strains with 100% primer match.
| Mycoplasmas | |||
|---|---|---|---|
| M. agalactiae | M. crocodyli | M. lagogenitalium | M. pulmonis |
| M. alligatoris | M. cynos | M. microti | M. salivarium |
| M. alvi | M. dispar | M. moatsii | M. sualvi |
| M. amphoriforme | M. edwardii | M. molare | M. synoviae |
| M. arginini | M. felis | M. mucosicanis | M. testudineum |
| M. bovigenitalium | M. fermentans | M. muris | M. testudinis |
| M. bovis | M. gallisepticum | M. mustelae | M. verecundum |
| M. buccale | M. genitalium | M. mycoides | M. volis |
| M. canadense | M. hominis | M. orale | M. yeatsii |
| M. canis | M. hyopneumoniae | M. oxoniensis | M. zalophi |
| M. capricolum | M. hyorhinis | M. penetrans | M. zalophidermidis |
| M. citelli | M. hyosynoviae | M. phocidae | |
| M. columborale | M. imitans | M. phocicerebrale | |
| M. conjunctivae | M. iowae | M. pirum | |
| M. cricetuli | M. lacerti | M. pneumoniae | |
| Ureaplasma | |||
| U. canigenitalium | U. diversum | U. gallorale | U. parvum |
| U. urealyticum | |||
| Mesoplasma | |||
| M. chauliocola | M. florum | M. photuris | M. tabanidae |
| M. entomophilum | M. grammopterae | M. syrphidae | |
| Spiroplasma | |||
| S. cantharicola | S. lineolae | S. taiwanense | S. citri |
| S. platyhelix | |||
| Acholeplasma | |||
| A. laidlawii |
|
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For more information, take a look at Biontex's Detection of Mycoplasma in Cell Culture FAQ here.