epicentre Cap Analog Trimethylated C06005 5 U

　　產(chǎn)品名稱:Cap Analog Trimethylated貨號(hào):C06005規(guī)格:5 U存儲(chǔ)溫度:-20℃

　　epicentre Cap Analog Trimethylated C06005 5 U產(chǎn)品描述Most naturally occurring eukaryotic mRNAs and many viral RNAs have a modified nucleotide structure known as a“cap”on their 5′terminus. In vivo, the cap structure is involved in the initiation of protein synthesis and in mRNA processing and stability.1 Many viral RNAs are infectious only when capped, and uncapped RNAs introduced into cells via transfection or microinjection are rapidly degraded by cellular RNases.2,3 Therefore,RNA repared for microinjection intooocytes or for transfection into eukaryotic cells should be capped. In vitro, capped mRNAs are translated more efficiently than uncapped transcripts in rabbit reticulocyte lysate or wheat germ translation systems.4 The trimethylated cap is involved in RNA transport5,6 and RNA translation in nematodes.7 Transcripts synthesized for use in in vitro RNA splicing experiments may also need to be capped.8 RNA Cap Analogs Cat. Nos. C32010, C61025, C06005, and C50110 Anti-reverse cap analogs (ARCAs)*, such as m2 7,3′–OG[5′]ppp[5′]G, were found to result in synthesis of transcripts that were more efficiently translated in vitro since the 3′-O-methyl group only permitted them to be incorporated in the correct orientation9,10,11 (see Figure 1). Capping does reduce RNA yields relative to transcription reactions without diguanosine triphosphate, but with the right conditions and high-quality reagents, a large proportion of the transcripts that are made will be capped. For optimum yields of monomethylated, capped in vitro transcripts, we recommend using the AmpliCap-Max?T7 and T3 High Yield Message Maker Kits. EPICENTRE also offers the MessageMAX?T7 ARCA-Capped Message Transcription Kit which is optimized to produce the highest yield of RNA having an ARCA on the 5′end. These kits contain a convenient Cap/NTP PreMix and yield up to 60μg of RNA perμg of Control Template in a standard 30-minute reaction. Capping efficiencies are up to 80%. EPICENTRE’s RNA Cap Analog preparations can also be used with the AmpliScribe?T7, T3, and SP6 High Yield Transcription Kits. Capping reactions with these kits typically yield about 20-40μg of total RNA with ~80% capping efficiency. Protocols for the synthesis ofcapped RNA using AmpliScribe or conventional transcription reagents are given on page 3.

　　epicentre Cap Analog Trimethylated C06005 5 U產(chǎn)品描述大多數(shù)天然存在的真核mRNA和許多病毒RNA在其5'末端具有修飾的核苷酸結(jié)構(gòu)，稱為“帽”。在體內(nèi)，帽結(jié)構(gòu)參與蛋白質(zhì)合成的啟動(dòng)以及mRNA的加工和穩(wěn)定性。1許多病毒RNA僅在加帽后才具有感染力，而通過(guò)轉(zhuǎn)染或顯微注射引入細(xì)胞的未蓋帽的RNA會(huì)被細(xì)胞RNase快速降解。2，3因此，用于顯微注射入卵母細(xì)胞或轉(zhuǎn)染到真核細(xì)胞中的RNA應(yīng)當(dāng)加蓋。在體外，在兔網(wǎng)織紅細(xì)胞裂解液或小麥胚芽翻譯系統(tǒng)中，加帽的mRNA比未加帽的轉(zhuǎn)錄本更有效地翻譯。4三甲基化帽參與線蟲中RNA的轉(zhuǎn)運(yùn)5、6和RNA的翻譯。7合成用于體外RNA剪接實(shí)驗(yàn)的轉(zhuǎn)錄本也可能需要加帽。8RNA Cap類似物Cat。發(fā)現(xiàn)編號(hào)為C32010，C61025，C06005和C50110的反反向帽類似物(ARCAs)*，例如m2 7,3'-OG [5'] ppp [5'] G，可以合成由于3'-O-甲基僅允許它們以正確的方向摻入9、10、11(請(qǐng)參見(jiàn)圖1)，因此在體外翻譯效率更高。相對(duì)于沒(méi)有三磷酸雙鳥苷的轉(zhuǎn)錄反應(yīng)而言，加帽的確降低了RNA的產(chǎn)量，但是在適當(dāng)?shù)臈l件和高質(zhì)量的試劑的條件下，大部分制成的轉(zhuǎn)錄本都將被加帽。為了獲得單甲基化，封端的體外轉(zhuǎn)錄本的最佳產(chǎn)率，我們建議使用AmpliCap-Max?T7和T3高產(chǎn)率消息產(chǎn)生試劑盒。EPICENTER還提供MessageMAX?T7 ARCA封頂?shù)南⑥D(zhuǎn)錄試劑盒，該試劑盒經(jīng)過(guò)優(yōu)化，可產(chǎn)生最高產(chǎn)量的5'末端帶有ARCA的RNA。這些試劑盒包含方便的Cap / NTP PreMix，在標(biāo)準(zhǔn)的30分鐘反應(yīng)中，每微克對(duì)照模板可產(chǎn)生高達(dá)60微克RNA。封蓋效率高達(dá)80%。EPICENTRE的RNA帽類似物制劑也可與AmpliScribe?T7，T3和SP6高產(chǎn)量轉(zhuǎn)錄試劑盒一起使用。這些試劑盒的加帽反應(yīng)通?？僧a(chǎn)生約20-40μg的總RNA，加帽效率約為80%。合成協(xié)議封蓋效率高達(dá)80%。EPICENTRE的RNA帽類似物制劑也可與AmpliScribe?T7，T3和SP6高產(chǎn)量轉(zhuǎn)錄試劑盒一起使用。這些試劑盒的加帽反應(yīng)通常可產(chǎn)生約20-40μg的總RNA，加帽效率約為80%。合成協(xié)議封蓋效率高達(dá)80%。EPICENTRE的RNA帽類似物制劑也可與AmpliScribe?T7，T3和SP6高產(chǎn)量轉(zhuǎn)錄試劑盒一起使用。這些試劑盒的加帽反應(yīng)通?？僧a(chǎn)生約20-40μg的總RNA，加帽效率約為80%。合成協(xié)議第3頁(yè)提供了使用AmpliScribe或常規(guī)轉(zhuǎn)錄試劑封端的RNA。

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