RNA 5’ Polyphosphatase
Product Description:
RNA 5' Polyphosphatase is a patented enzyme that catalyzes the
removal of γ and β phosphate groups from 5'-triphosphorylated and
5'-diphosphorylated RNA resulting in 5'-monophosphorylated RNA. Capped
RNA is not a substrate for the enzyme.
RNA 5' Polyphosphatase can be used for the following applications:
- Removal of uncapped RNA from an mRNA preparation, in conjunction with XRN1 treatment.
- 5' end RNA substrate preparation for RNA Ligase-based circularization reactions.
- 5' end RNA substrate preparation for T4 RNA Ligase-based reactions (end tagging).
RNA 5' Polyphosphatase is active on nucleoside triphosphates and is
inhibited by inorganic phosphate, a natural reaction product. Removal of
unincorporated nucleotides and inorganic phosphates from completed
transcription reactions is recommended for full enzymatic activity on
the intended RNA substrate.
- Catalyzes the removal of γ and β phosphate groups from
5'-triphosphorylated and 5'- diphosphorylated RNA, resulting in
5’-monophosphorylated RNA. - Two protocols provided for easy- and hard-to-treat samples.
- Use in conjunction with XRN1 to remove uncapped RNA from
mRNA preparations. - Use for 5’ end RNA substrate preparation for
T4 RNA Ligase-based reactions.
Product is for research use only (RUO)
Product performance
Figure 1. Two different 5'-triphosphate primary
transcript RNAs were treated with RNA 5' Polyphosphatase using the
Standard Protocol (no pre heat denaturation step used). After the 30
minute incubation, additional MgCl2 (for buffer
compatibility) and XRN1 (5'-monophosphate-dependent exonuclease) was
added to the reaction to digest the RNA 5' Polyphosphatase reaction
product. Following treatment with RNA 5’ Polyphosphatase and XRN1, the
1.85-kb RNA (lane 2) and 0.92-kb RNA (lane 4) were digested.
Figure 2. A 1.57-kb 5'-triphosphate primary transcript
RNA containing stable secondary structure on the 5' end was treated with
RNA 5' Polyphosphatase using the Alternate Protocol (pre heat
denaturation step used). After the 30 minute incubation, additional
MgCl2 (for buffer compatibility) and XRN1
(5'-monophosphate-dependent exonuclease) was added to the reaction to
digest the RNA 5' Polyphosphatase reaction product. Heat denaturation
prior to RNA 5’ Polyphosphatase and XRN1 treatment improved reaction
efficiency (lane 6).
| Components |
Volume |
RNA 5' Polyphosphatase, 20 U/µl
in 50% glycerol, 50 mM Tris-HCl, pH 7.5, 0.1 M NaCl, 0.1 mM EDTA and 0.1% Triton® X-100. |
20 µl |
10X RNA 5' Polyphosphatase Reaction Buffer
0.5 M HEPES-KOH, pH 7.5, 1 M NaCl, 10 mM EDTA, 1% β-mercaptoethanol, and 0.1% Triton X-100. |
100 µl |
Manual
SDS
Product Flyer
If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200