This product is a polymerase expressed by recombinant E.coli. The enzyme does not depend on the template and can catalyze the sequential incorporation of ATP into the 3′ end of RNA in the form of AMP, that is, adding a polyadenosine tail to the 3′ end of RNA. Poly(A) structure can improve the stability of RNA and improve the translation efficiency of mRNA in eukaryotic cells. Poly(A) polymerase has high tailing efficiency and can add 20-200 A bases to the 3′ end of RNA.
Storage and transportation
Store at -20 ℃, Transportation by dry ice.
Unit Definition
One unit is defined as the amount of enzyme that will incorporate 1 nmol of AMP into RNA in a 20 µl volume in 10 minutes at 37°C.
Quality Control
Purity≥95%, no DNase, RNase activity, host DNA residue≤100pg/mg, host protein residue≤50ppm, endotoxin residue≤10EU/mg, no protease activity, sterile, no mycoplasma.
Reaction System and Condition
| Component Name | Amount |
|---|---|
| 10 × Poly(A) Polymerase Buffer | 2 μL |
| E.coli Poly(A) Polymerase (5 U/μl) | 0.2 – 2 μl |
| ATP (10 mM) | 1 μl |
| Denatured Cap0 RNA | 10 μg |
| RNase-free ddH2O | Up to 20 μL |
Notes
- The length of the Poly(A) tail added in this reaction is affected by factors such as enzyme amount, ATP concentration and reaction time. Generally, this enzyme can add ~30 A bases for 30min at 37℃, and ~100 A bases in 60min. .
- The enzyme adds AMP to the 3′ end of RNA with high selectivity, and does not add the same length of Poly(A) to all RNA molecules.
- The substrate of this enzyme can only be RNA, and it needs divalent ions such as Mg2+ to be active.
- This enzyme can also be reacted with M-MuLV Reverse Transcriptase Reaction Buffer
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