- 询价
- Hongene
- 上海
- ON-126
- 2025年07月03日
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- 技术资料
- 保存条件：
-20℃
- 保质期：
N/A
- 英文名：
Poly(A) Polymerase
- 库存：
N/A
- 供应商：
Hongene/上海兆维
- 规格：
＞95% by SDS-PAGE
产品描述：Poly(A) polymerase是 以ATP为底物对RNA的3'羟基端添加腺苷酸形成polyA尾部结构。
来源：重组大肠杆菌
规格：5U/μL
活性单位定义：1单位活性定义为20μL的反应体系，37℃、10分钟内，掺入1nmol AMP 到酸性不溶物质所需的酶量。
酶存储缓冲液：20mM Tris-HCl (pH7.5, 25℃), 1mM EDTA, 300mM NaCl, 1mM DTT, 0.1%(v/v) Triton X-100, 50%(v/v) glycerol
配套溶液：10X poly(A) Polymerase Reaction Buffer, Cat#ON-127, 500mM Tris-HCl, (pH7.9, 25℃), 2.5M NaCl, 100mM MgCl2
相关产品推荐：10X poly(A) Polymerase Reaction Buffer, Cat#ON-127; Taq DNA Polymerase, Cat#ON-007; RNase inhibitor, Cat#ON-039; pfu DNA polymerase, Cat#ON-051; High Yield T7 RNA Synthesis Kit, Cat#ON-040
操作要点：
操作流程：本流程适用于小于10μg的RNA添加150bp长度的poly(A)尾部结构。来源于IVT反应的RNA需要纯化后，才能用于poly(A)加尾反应。
1. 按下列配方配置反应液：
| RNA | 1~10μg |
| 10X poly(A) Polymerase Reaction Buffer | 2μL |
| ATP(10mM) | 2μL |
| Poly(A) polymerase | 1μL |
| (可选) Rnase inhibitor | 20units |
| Nuclease-free H2O | to 20μL |
2. 37度保温60分钟
3. 终止反应，加入EDTA至终浓度大于11mM
* 热变性的RNA可能会提高加尾反应的效率
储藏温度：-20℃
质控检测：无DNase、Nickase、RNase污染
纯度：SDS-PAGE纯度：大于95%
参考文献：
1. Bernstein P and Ross J (1989) Poly(A), poly(A) binding protein and the regulation of mRNA stability. Trends Biochem Sci 14: 373–377.
2. Gallie DR (1991) The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency. Genes Dev 5: 2108–2116.
3. Krug, M.S. and Berger, S.L. (1987) Methods Enzymol. 152, 262.
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