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產(chǎn)品信息

     The Poly(A) Polymerase Tailing Kit was developed for the rapid and efficient addition of poly(A)-tails to  the 3′-hydroxyl end of any RNA. RNA with 3′ phosphates may be  endrepaired using T4 Polynucleotide Kinase (PNK) in the absence of ATP. A simple cleanup will be necessary to remove T4 PNK prior to A-tailing.Polyadenylation increases the stability of RNA in eukaryotic cells and enhances its ability to be translated after transfection or microinjection.Poly(A) tails can also provide priming sites for the synthesis of first-strand cDNA, or be used to end-label4 or quantitate5 mRNA.   
The kit features Poly(A) Polymerase which uses ATP as a substrate for template-independent addition of adenosine monophosphates to the 3′-hydroxyl termini of RNA molecules. The standard protocol was designed to produce a poly(A)-tail length of ~150 b on 60 μg of capped RNA.An alternative protocol is also provided for tailing lesser amounts of RNA as well as suggestions on how to adjust the length of the poly(A)-tails generated.
      Poly（A）聚合酶尾巴試劑盒是為快速有效地將poly（A）尾添加到任何RNA的3'-羥基末端而開發(fā)的。 在不存在ATP的情況下，可以使用T4多核苷酸激酶（PNK）對(duì)帶有3'磷酸的RNA進(jìn)行末端修復(fù)。在A尾之前，必須先進(jìn)行簡單的清除以去除T4 PNK.Polyadenylation可提高真核細(xì)胞中RNA的穩(wěn)定性并增強(qiáng)其在轉(zhuǎn)染或顯微注射后的翻譯能力.Poly（A）尾部還可為合成提供啟動(dòng)位點(diǎn) 第一鏈cDNA的序列，或用于末端標(biāo)記4或定量5 mRNA。
    該試劑盒具有Poly（A）聚合酶，該酶使用ATP作為底物，將模板磷酸單磷酸腺苷獨(dú)立地添加到RNA分子的3'-羥基末端。 標(biāo)準(zhǔn)操作設(shè)計(jì)為在60μg帶帽RNA上產(chǎn)生?150 b的poly（A）尾長。還提供了一種替代方案，用于拖尾較少量的RNA，以及有關(guān)如何調(diào)整生成的poly（A）尾巴長度的建議。
    Poly(A)聚合使用ATP作為底物，用于模板依賴性地將腺苷一磷酸加成到RNA分子的3-OH末端。Poly(A)聚合酶加尾試劑盒提供酶和其他試劑，可快速、輕松地將poly(A)尾部添加到任何RNA的3’末端。

優(yōu)點(diǎn)：
? Quality Kit: High enzymatic purity, specificity, and activity
? Flexible: Use polyadenylated RNAs in a variety of applications such as stabilizing in   vitro transcribed RNA and adding primer (oligo-dT) binding sites to any RNA  for first-strand cDNA synthesis
? Fast and Easy: Simple protocol produces polyadenylated RNA molecules quickly
圖示

Figure 1. Customized Poly(A)-tail Lengths. A 1.4-kb transcript was poly(A)-tailed using various reaction conditions to demonstrate the affect on poly(A)-tail   lengths. Each lane contains 0.1 μg of the completed poly(A)-tailing reaction product.
圖1.定制的Poly（A）尾部長度。 使用各種反應(yīng)條件將1.4-kb的轉(zhuǎn)錄本進(jìn)行聚（A）尾部修飾，以證明其對(duì)聚（A）尾部長度的影響。 每個(gè)泳道包含0.1μg完整的poly（A）尾反應(yīng)產(chǎn)物。
組成成分：
? Poly(A) Polymerase (4 U/μL)：儲(chǔ)存在-20°C
? ATP（10mM）：儲(chǔ)存在-20°C
? 10X Poly(A) Polymerase Reaction Buffer：儲(chǔ)存在-20°C
? Nuclease-Free Water：儲(chǔ)存在-20°C
參考文獻(xiàn)：
1. Drummond, D.R. et al., (1985) J. Cell. Biol. 100, 1148.
2. Galili, G. et al., (1988) J. Biol. Chem. 263, 5764.
3. Belasco, J. and Brawerman, G. (1993) Control of Messenger RNA Stability,   Academic Press, San Diego, CA.
4. Lingner, J. and Keller, W. (1993) Nucleic Acids Res. 21, 2917.
5. Krug, M.S. and Berger, S.L. (1987) Methods Enzymol. 152, 262.