Cell Culture and Transfection
2X MycoPCR Master Mix is a 2X premixed, ready-to-use solution (GeneSpin proprietary formulation) containing XtraTaq Pol, dNTPs, MgCl2, specific mycoplasma primers set and stabilizers optimized for PCR-based mycoplasma detection in cell cultures. 2X MycoPCR Master Mix contains a combination of tartrazine and xylene dyes that allow gel loading and electrophoresis of the sample directly from the PCR tube, without further manipulation.
The primer set allows detection of various mycoplasma species (e.g., M. fermentans, M. hyorhinis, M. arginini, M. orale, M. salivarium, M. hominis, M. pulmonis, M. arthritidis, M. bovis, M. pneumoniae, M. pirum and M. capricolum), as well as Acholeplasma and Spiroplasma species, with high sensitivity and specificity.
6x25 assay kit
2x25 assay kit
25 assay kit
FOR RESEARCH USE ONLY
Shipped in green ice.
Store at 4C°
1. Transfer 100-200ul of cell culture supernatant into 1.5ml centrifuge tube. Incubate the supernatant at 95°C for 5 minutes.
2. Centrifuge at maximum speed for 5 minutes.
3.Use 2-5ul of the supernatant as PCR template.
IMPORTANT! Before harvesting the supernatant from the cell culture, cells should cover approximately 90% of the growth surface! The supernatant may cause PCR inhibition in excessively dense cell cultures (>90%).
4. For optimum separation we recommend using a 2% agarose gel with TAE or TBE buffer used for electrophoresis.
5. Load all PCR product volume (20ul) directly onto the gel and perform electrophoresis.
6. When the electrophoretic run is completed, lay the gel onto UV transilluminator to detect the expected band.
7. IF the test is POSITIVE, a DNA band of 750bp will appear.
PCR assay set-up:
2X Muco PCR Mix
template from step 3
Spin down the tubes/plate briefly to remove bubbles and place them into the cycler.
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