Applichem A3744 支原体PCR检测试剂盒 PCR Mycoplasma Test Kit

【简单介绍】
AppliChem推出的PCR Mycoplasma Kit（货号：A3744）是一个即用型支原体检测试剂盒，无需支原体培养，只需要从培养细胞液中取上清，离心后获得可能含有支原体的沉淀，用50ul Kit内提供的缓冲液重悬后，95℃热击3分钟后即可作为PCR模板，加入Kit内提供的PCR反应液，并配成50ul PCR反应体系后即可按照Protocal提供的PCR设置进行PCR反应（如左图所示）。同时该支原体检测Kit内提供阳性对照，可完成20次检测。

Product Code:  A3744

Product Name:  PCR Mycoplasma Test Kit

Specifications:

♦ Ready-to-use PCR Mix for the detection of mycoplasma in cell culture
♦ detects all of myoplasma species found in cell cultures
♦ for 20 tests
♦ Kit components
• Reaction mix
• Buffer solution
• Positive template control
• Internal control DNA template
• Internal control primers mix

Comment:

The PCR Mycoplasma Test Kit is designed to detect the presence of mycoplasma contaminating biological materials, such as cultured cells. Mycoplasma detection by the direct culture procedure is time-consuming and some mycoplasma species are difficult to cultivate. With PCR testing, results are obtained within a few hours, since the presence of contaminant mycoplasma can be easily detected simply by verifying the bands of amplified DNA fragments in electrophoresis. There is no need to prepare probes labeled with radioisotopes, or to calculate enzyme, dNTP’s or buffer concentrations. Instead, a ready-to-use, optimized PCR mix is supplied. The primer set allows detection of various mycoplasma species (M. fermentans, M. hyorhinis, M. arginini, M. orale, M. salivarium, M. hominis, M. pulmonis, M. arthritidis, M. bovis, M. pneumoniae, M. pirum, M. capricolum) as well as Acholeplasma and Spiroplasma species, with high sensitivity and specificity.Principle\: rRNA gene sequences of prokaryotes, including mycoplasmas, are well conserved, whereas, the lengths and sequences of the spacer region in the rRNA operon (for example the region between 16S and 23S gene) differ from species to species. The detection procedure utilizing the PCR process with this primer set consists of1.) Amplification of a conserved and mycoplasma-specific 16S rRNA gene region using two primers.2.) Detection of the amplified fragment by agarose gel electrophoresis.This system does not allow the amplification of DNA originating from other sources, such as cultured cells or bacteria, which affect the detection result. Amplification of the gene sequence with PCR using this primer set enhances not only the sensitivity, but also the specificity of detection. Amplified products are then detected by agarose gel electrophoresis.

Bibliography:

(1)Rottem, S. & Barile, F.M. (1993) TIBTECH 11, 143-150

Storage:  -20°C
Avoid repeated changes in the reaction mix temperature
When in use, always keep the reaction mix on ice!

Shipment:   wet ice in Germany, dry ice to abroad

【详细说明】

针对支原体检测，有多种方案可供选择，包括直接培养法、DNA荧光染色法等。其实就支原体污染的检测，zui简单有效的方案不外乎是使用PCR方法检测。即以支原体16S-23S之间极为保守的rRNA序列设计引物，经由PCR反应扩增其16S rRNA片段。该法非常灵敏（e.g. 0.1-1.6 CFU/5 ul 样品） 。由于引物的特殊性，不会导致培养细胞或其他细菌的序列的假性扩增。方法便捷，只需不到一天时间，并且无需支原体或细胞的培养过程，避免了可能的污染。