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Vaccinia Capping Enzyme Vaccinia Capping System 牛痘病毒加帽酶 牛痘病毒加帽体系 替代NEB M2080S NEB M2081S NEB M2081L
Analysis of mRNA capping efficiency using LC-MS
真核生物中，mRNA经转录后修饰在5'端形成一个特殊的帽子结构(m7Gppp5'N)，此结构与mRNA的稳定性、转运(出核)和翻译紧密相关。通过酶促反应为RNA加帽是一种简单有效的方法，能够显著改善体外转录的用于转染和显微注射的RNA的稳定性和翻译能力。牛痘病毒加帽酶(Vaccinia Capping Enzyme)由两个亚基(D1，D12)组成，兼具RNA三磷酸酯酶活性、鸟苷酰基转移酶活性和鸟嘌呤甲基转移酶活性，它们对于添加一个完全的Cap0结构都是必须的。在合适浓度的S-腺苷甲硫氨酸(S-Adenosylmethionine，SAM)，鸟苷三磷酸(GTP)等条件存在下，牛痘病毒加帽酶能够将帽子结构(m7Gppp，Cap0)加到RNA5'端。基于本酶促反应，加帽效率可接近100%，并且保证帽子结构方向正确。
本制品可用于T7 RNA Polymerase催化的体外转录反应产生的RNA的加帽反应。
Product Information
| Source | E. coli carrying the capping enzyme gene of vaccinia virus |
Storage Buffer | 20mM Tris-HCl, pH 8.0 |
Transportation/ Storage Method | Transportation and storage at 20 ± 5 ℃, in order to avoid repeated freezing and thawing, it is recommended to repack immediately after receiving |
Enzyme Activity Definition | One unit (1U): The amount of enzyme required to incorporate 10 pmol (α-32P) GTP into an 80-nucleotide (80nt) transcript in 1 hour at 37°C |
Product application | 1, Capping reaction of pre-mRNA translation in vivo/in vitro. |
Product Components
| Components | Cat. No. / Size | |
VCS-DL101-B(500U) | VCS-DL101-C(5000U) | |
Vaccinia Capping Enzyme (10U/μl) | VCS-DL101-B1(50μl) | VCS-DL101-C1 (500μl) |
10 ×Capping Buffer | VCS-DL101-B2(200μl) | VCS-DL101-C2 (1.5ml) |
GTP(10mM) | VCS-DL101-B3(50μl) | VCS-DL101-C3(500μl) |
SAM(32mM) | VCS-DL101-B4(50μl) | VCS-DL101-C4(500μl) |
Other Notes
1) The RNA used for the capping reaction must be purified and resuspended in RNase-free Water.
2) Heat treatment of RNA before the reaction to remove secondary structure at the 5' end.
3) If the structure of the RNA 5' end is known, the reaction time can be extended to 60min to improve the capping efficiency.
4) In the 5' end labeling reaction, the GTP stock solution should be diluted to 1-3 times the molar concentration of mRNA in the reaction system.
5) It is recommended to use RNase inhibitors to enhance the stability of RNA in the reaction. Add 0.5μl of RNase inhibitor (eg Murine RNase Inhibitor, RNI-DL001) during the reaction.
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