Detection primers, kit and detection method for cell mcoplasma

Abstract

The invention belongs to the technical field of biology and particularly relates to detection primers, a kit and a detection method for cell mcoplasma. The kit comprises a combination of two pairs ofprimers required for PCR (Polymerase Chain Reaction) amplification, wherein the primer combination can specifically amplify approximate 250bp sequences between 16S genes and 23S genes in various mycoplasmas; meanwhile, the amplification of two pairs of primers ensures the accuracy of test results. The kit also comprises a pre-reactive mixture required for the PCR amplification; the mixture comprises Taq enzymes, dNTP, a buffer solution and the like; the kit disclosed by the invention can quickly and efficiently distinguish whether mycoplasma contamination in the cell occurs or not and can obtain accurate detection result; meanwhile, batch detection can be realized.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a detection primer, a kit and a detection method for mycoplasma cells. Background technique [0002] Mycoplasma is a prokaryotic microorganism with a size between bacteria and viruses (0.1 - 0.6 μm) and no cell wall. Because the size of mycoplasma is very small, about 1% of mycoplasma can pass through the filter membrane with a pore size of 0.2 μm in the laboratory. At the same time, mycoplasma can easily reproduce in artificial medium culture. Mycoplasma pollution has become a major source of pollution in cell culture. Most mycoplasma are not sensitive to common antibiotics, so it is difficult to kill mycoplasma, and the cell state after light pollution has no change whether it is observed with the naked eye or under a common microscope. Therefore, it is easy to ignore the problem of mycoplasma pollution. Existing reports have shown that the proportion of mycoplasma ...

AI Technical Summary

Problems solved by technology

Some PCR detection methods that have been published at present mainly carry out PCR amplification through the conservative synthetic primers of mycoplasma rRNA sequence, but these methods have high requirements on the experimental environment, and the experimental cost is relatively expensive, and the most important thing is that false positives are prone to occur. limited

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