Applichem A3744 支原體PCR檢測試劑盒 PCR Mycoplasma Test Kit

【簡單介紹】
AppliChem推出的PCR Mycoplasma Kit（貨號(hào)：A3744）是一個(gè)即用型支原體檢測試劑盒，無需支原體培養(yǎng)，只需要從培養(yǎng)細(xì)胞液中取上清，離心后獲得可能含有支原體的沉淀，用50ul Kit內(nèi)提供的緩沖液重懸后，95℃熱擊3分鐘后即可作為PCR模板，加入Kit內(nèi)提供的PCR反應(yīng)液，并配成50ul PCR反應(yīng)體系后即可按照Protocal提供的PCR設(shè)置進(jìn)行PCR反應(yīng)（如左圖所示）。同時(shí)該支原體檢測Kit內(nèi)提供陽性對照，可完成20次檢測。

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

【詳細(xì)說明】

針對支原體檢測，有多種方案可供選擇，包括直接培養(yǎng)法、DNA熒光染色法等。其實(shí)就支原體污染的檢測，zui簡單有效的方案不外乎是使用PCR方法檢測。即以支原體16S-23S之間極為保守的rRNA序列設(shè)計(jì)引物，經(jīng)由PCR反應(yīng)擴(kuò)增其16S rRNA片段。該法非常靈敏（e.g. 0.1-1.6 CFU/5 ul 樣品） 。由于引物的特殊性，不會(huì)導(dǎo)致培養(yǎng)細(xì)胞或其他細(xì)菌的序列的假性擴(kuò)增。方法便捷，只需不到一天時(shí)間，并且無需支原體或細(xì)胞的培養(yǎng)過程，避免了可能的污染。