SKU : G1902-50
Categories : 2. Cell & Molecular Biology , Cell Culture Medium / Sera , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat.No. | Spec. |
One-step Rapid Mycoplasma Detection Kit (Isothermal Amplification Method) | G1902-50 | 50 rxns |
G1902-100 | 100 rxns |
Description/Introduction
The kit can quickly detect eight kinds of mycoplasma in cell culture medium by isothermal amplification technique and visual color change. Without DNA extraction, 1 μL cell culture supernatant was directly added to the reaction solution and 30 min was reacted at 65°C. The color of the positive sample changed from yellow to red, and the whole process does not need PCR instrument or open lid to electrophoresis, which can effectively avoid the cross contamination caused by aerosol. Compared with the traditional PCR method, this product has the advantages of simple operation, high sensitivity, and can tolerate a variety of PCR inhibitors in the culture medium, so there is no false negative phenomenon, and the detection results are highly consistent with qPCR.
Storage and Handling Conditions
Shipped with wet ice and stored at -20°C; valid for up to 12 months.
Product Contents
Component Number | Component | G1902-50 | G1902-100 |
1902-1 | Mycored Buffer | 1.2 mL | 1.2 mL×2 |
1902-2 | Mycored Enzyme | 50 µL | 100 µL |
1902-3 | Positive Control | 50 µL | 100 µL |
1902-4 | Paraffin Oil | 1 mL | 1 mL×2 |
Manual | One copy | ||
a：Contains chromogenic agents.
Before starting (please read carefully)
Set up 65°C water bath or metal heating block in advance.
Assay Protocol / Procedures
1. Sample preparation to be tested:
a. Adherent cells: Absorb the supernatant directly. It is suggested that the samples should be taken when the cell inoculation or liquid change is more than 3 days and the confluence degree is about 90%. At this time, the content of mycoplasma in the supernatant is high and can be easily detected.
b. Suspension cells: The supernatant was obtained after centrifugation at 1,000 rpm for 5 min. It is suggested that the samples should be taken after cell inoculation or fluid change for more than 3 days, when the content of mycoplasma in the cell culture medium is high and can be easily detected.
2. Reaction system
Component | 25 μL |
Mycored Buffer | 23 μL |
MycoRed Enzyme | 1 μL |
a: Mycored Buffer is used upside down after thawing.
b: According to the number of samples, after the above reaction system is prepared, sub-pack it into the reaction tube, add 20 μL of Paraffin Oil, and then proceed to the next step.
3. Add samples
a. Negative control: No sample was added to the first reaction tube as a negative control.
b. Positive control: 1 μL Positive Control was added to the last reaction tube as positive control.
c. Sample to be tested: Add 1 μL cell culture supernatant to the rest of the reaction tube.
a: Please extend the head of the pipette tips under the liquid level of paraffin oill to add samples.
4. Reaction condition
Transfer the reaction tube to a preheated 65 water bath or metal bath to incubate 30min. If the color development is not obvious the reaction time can be extended to 40 min.
Result determination
Take out the reaction tube immediately after the end of the reaction, and if the reaction solution is still yellow after it is restored to room temperature, the result will be negative. If the reaction solution turns red, the result is positive. The reaction tube should not be opened, otherwise it will easily lead to aerosol pollution.
Note
1. Do not open the reaction tube after the reaction is over.
2. It is suggested that the pipette tips with filter element should be used in the process of preparing reaction solution, and the discarded pipette tips and reaction tube should be put into the self-sealing bag and dealt with in time.
3. Please use nucleic acid scavenger (recommended G3020) to wipe the test bench and pipette frequently.
For Research Use Only!
