Mycoplasma are common and serious contaminants of cell cultures. It has been shown that 30% to 87% of cell cultures are infected with mycoplasma. Many mycoplasma contaminations, particularly in continuous cell lines, grow slowly and do not destroy host cells but are still able to affect various parameters, leading to unreliable or false results. These effects include changes in metabolism, growth, viability, DNA, RNA, and protein synthesis, and morphology. Testing for mycoplasma is an essential quality control tool to assure accurate research and reliable biotechnological products. The e-Myco™ (ver.2.0) product is a set of primers designed to detect the presence of mycoplasma that might contaminate biological materials such as cultured cells. Conventional techniques that are used to detect mycoplasma involve culturing samples on selective media, which needs at least 1 week to obtain results, whereas by performing PCR with this primer set, which is based on conserved 16S rRNA, detection results are obtained in a few hours.
The adopted 8-methoxypsoralen (8-MOP) is used to extinguish the template activity of contaminated DNAs. 8-MOP is known to intercalate into double-stranded nucleic acids and form a covalent interstrand crosslink after photo-activation by incident light at wavelength 320-400 nm. An internal control of this product was constructed to identify false negative results in each reaction. The internal control was designed in such a way that the sample primer pair was used to amplify the internal control and target DNA, which were differentiated by size
■ Premix Type
- This e-Myco™ Mycoplasma PCR Detection Kit(ver.2.0) contains all the components.
■ Broad Species Detection
- You can detect five common cell culture-infecting species of mycoplasma and also other various species of mycoplasma (See Technical Guide).
■ Species Determination
- You can determine the species of mycoplasma by sequencing the amplified PCR products.
■ Internal Control (Instead of positive control)
- Internal control embedded in the product prevents misjudgment that possibly arises from an erroneous PCR test.
■ Elimination of Carryover Contamination System
- 8-MOP solution prevents carryover contamination by PCR products
■ PCR Reaction volume
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20 μl reaction |
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e-MycoTM Mycoplasma PCR Detection Kit (ver. 2.0)
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i-StarTaqTM DNA Polymerase
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2.5U
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Chemical Stabilizer
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1X
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Loading Buffer
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1X
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dNTPs
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250mM each
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Tris-HCl (pH 8.3)
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10mM
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| KCl |
50mM
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| MgCl2 |
1.5mM
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Mycoplasma Primer Sets
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10pmol/each
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| Internal Control |
| 8-MOP (dissolved in DMSO) |
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e-Myco™ Mycoplasma PCR Detection Kit (ver.2.0)
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100㎕(1U/㎕)
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| DNase/RNase-free Distilled Water |
1 vial (1 ml)
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Overview of Mycoplasma Detection
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