Vaccinia Virus Capping Enzyme HYJ231
Vaccinia virus capping enzyme, derived from a recombinant Escherichia coli strain carrying the vaccinia virus MR, consists of two subunits, D1R and D12L, and possesses three enzymatic activities: RNA triphosphatase, guanylyltransferase, and guanine methyltransferase. Vaccinia virus capping enzyme is an efficient enzyme that catalyzes the formation of a cap structure, specifically adding a 7-methylguanylate cap structure (m7Gppp, Cap 0) to the 5' end of RNA. The cap structure (Cap 0) plays a crucial role in the stability, transport, and translation of eukaryotic mRNA. Enzymatic RNA capping is an effective and simple method that can significantly improve the stability and translational capacity of RNA for in vitro transcription, transfection, and microinjection.
Components
|
Components |
2KU |
10KU |
100KU |
1000KU |
|
Vaccinia Virus Capping Enzyme (10 U/μL) |
0.2 mL |
1 mL |
10 mL |
100 mL |
|
10× Capping Buffer |
0.4 mL |
2×1mL |
2×10 mL |
2×100mL |
Storage
Store at -25 to -15°C. Valid for 2 years (avoid repeated freezing and thawing).
Storage buffer
20 mM Tris-HCl (pH 8.0), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.1% Triton X-100, 50% glycerol.
Activity Definition
One unit (U) of activity is defined as the amount of enzyme required to completely incorporate 10 pmol of GTP into an 80-nt transcript within 1 h at 37°C.
Quality Control
1.Exonuclease activity: When 10 U of this product was reacted with 1 μg of λ-Hind III digested DNA at 37°C for 16 hours, the electrophoretic band of the DNA remained unchanged.
2.Endonuclease activity: When 10 U of this product was reacted with 1 μg of λ DNA at 37°C for 16 hours, the electrophoretic band of the DNA remained unchanged.
3.Nickase activity: When 10 U of this product was reacted with 1 μg of pBR322 at 37°C for 16 hours, the electrophoretic band of the DNA remained unchanged.
4.RNase activity: When 10 U of this product was reacted with 1.6 μg of MS2 RNA at 37°C for 4 hours, the electrophoretic band of the RNA remained unchanged.
5.Residual E. coli DNA: Residual nucleic acid in 10 U of this product was detected by TaqMan qPCR specific for E. coli 16s rDNA, and the E. coli genome residue was ≤1. Copy;
6.Endotoxin: LAL-Test, Chinese Pharmacopoeia 2020 Edition Part IV Gel Limit Test General Rules(1143) , bacterial endotoxin content ≤ 10 EU/mg.