3. Mycoplasma Detection Kit (PCR); 50T $300

Description

Mycoplasma contamination will cause adverse effects on all aspects of cells, and has become a highly valued problem in cell culture. Therefore, it is very necessary to regularly detect mycoplasma contamination. This mycoplasma detection kit uses PCR method to specifically detect mycoplasma contamination in various cultured cell biological materials (such as cell cultures, secretions of experimental animals, animal serum, etc.). The mixed primers used in the kit are specific primers designed for the conserved region of the 16S rRNA sequence of Mycoplasma. The specific PCR amplification of Mycoplasma DNA can be completed within one hour with the matching Mycoplasma PCR Mix (2×) using cell culture solution as the PCR template. This kit has high sensitivity and can detect as little as 20 copies of mycoplasma.

Storage and Handling Conditions

Wet ice bag transportation; Stored at -20℃ temperature, valid for 12 months.

Component

Assay Protocol

1. Prepare the sample: Appropriate amount of cell culture supernatant to be tested was taken into a clean PCR tube. This PCR tubes was heat-treated at 95°C for 5 min in PCR instrument, and used it as a template. Serum samples could be diluted with Mycoplasma Free Water, and appropriate samples were placed in a clean PCR tube. This PCR tubes was heat-treated at 95°C for 5 min in PCR instrument, and used it as a template. ；

2. Prepare the PCR reactions: Negative control (replace 1 μL of sample with the same amount of Mycoplasma Free Water) and positive control (add 0.5 μL Positive Control to 1 μL sample as Template) shall be set for each experiment. Wear disposable masks and gloves during the experiment, and operate with caution to prevent improper introduction of exogenous mycoplasma contamination;

3. Setting up the PCR program:

4. Gel electrophoresis: Take 10 μL PCR product and use 1% agarose gel for electrophoresis detection.

5. Analyze the results: In each experiment, the Mycoplasma contamination of the samples by comparing with the results of negative control and positive control. The positive band size was about 500 bp. If there are bands in the negative control test results, it is likely to be contamination in the PCR system, and it is suggested to confirm the results again. If necessary, PCR products can also be routinely sequenced to identify specific mycoplasma species.

Note

1. This kit can detect M. orale，M. arginini，M. bovis，M. fermentans，M. gallisepticum，M. hominis，M. pirum，Ureaplasma spp，M. hyorhinis，M. pneumoniae，A. laidlawii and other at least 11 kinds of mycoplasma contamination.

2. All reagents should be thawed and mixed thoroughly on ice before using, and stored at -20℃ after used.

3. Negative controls and positive controls must be set for each experiment, and the experimental group recommends setting sample template gradients.

4. Masks must be worn during the operation, and PCR operation standards must be strictly followed to prevent the introduction of exogenous pollution from affecting the experimental results.

5. In order to ensure the reliability and stability of cell experiments, it is recommended that mycoplasma contamination be detected regularly.

6. For your safety and health, please wear lab coat and disposable gloves.

For Research Use Only!