产品信息

     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）聚合酶尾巴试剂盒是为快速有效地将poly（A）尾添加到任何RNA的3'-羟基末端而开发的。 在不存在ATP的情况下，可以使用T4多核苷酸激酶（PNK）对带有3'磷酸的RNA进行末端修复。在A尾之前，必须先进行简单的清除以去除T4 PNK.Polyadenylation可提高真核细胞中RNA的稳定性并增强其在转染或显微注射后的翻译能力.Poly（A）尾部还可为合成提供启动位点 第一链cDNA的序列，或用于末端标记4或定量5 mRNA。
    该试剂盒具有Poly（A）聚合酶，该酶使用ATP作为底物，将模板磷酸单磷酸腺苷独立地添加到RNA分子的3'-羟基末端。 标准操作设计为在60μg带帽RNA上产生〜150 b的poly（A）尾长。还提供了一种替代方案，用于拖尾较少量的RNA，以及有关如何调整生成的poly（A）尾巴长度的建议。
    Poly(A)聚合使用ATP作为底物，用于模板依赖性地将腺苷一磷酸加成到RNA分子的3-OH末端。Poly(A)聚合酶加尾试剂盒提供酶和其他试剂，可快速、轻松地将poly(A)尾部添加到任何RNA的3’末端。

优点：
▪ 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
图示

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.
图1.定制的Poly（A）尾部长度。 使用各种反应条件将1.4-kb的转录本进行聚（A）尾部修饰，以证明其对聚（A）尾部长度的影响。 每个泳道包含0.1μg完整的poly（A）尾反应产物。
组成成分：
▪ Poly(A) Polymerase (4 U/μL)：储存在-20°C
▪ ATP（10mM）：储存在-20°C
▪ 10X Poly(A) Polymerase Reaction Buffer：储存在-20°C
▪ Nuclease-Free Water：储存在-20°C
参考文献：
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.