Product Overview
Description
Ribonuclease Inhibitor
RNase Inhibitor is a recombinant protein isolated from E. coli strain carrying human placenta
ribonuclease gene. It specifically inhibits RNase A, RNase B, RNase C and forms 1:1 complex
with RNase. It is unable to inhibit RNase 1, RNase T1, S1 nuclease RNase H and Aspergillus-
-originated RNase. It shows no inhibitory effect on DNA Polymerases, AMV, M-MLV, SP6,
T7 and T3 RNA Polymerases.
| Cat. No. | AI101 | |
| Specification | AI101-01 | 2000 units |
| AI101-02 | 5×2000 units | |
| Concentration: | 50 units/μl | |
| Storage: | at -20°C for two years | |
| Shipping | Dry ice (-70 ℃) | |
| Storage Buffers | 50 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 10 mM DTT, 100 mM NaCl, 50% glycerol | |
| Unit Definition | One unit (U) is defined as the amount of the polymerase required for inhibiting 50% of 5 ng RNaseA activity. | |
| Applications |
cDNA synthesis, polysome extraction, in vitro translation, in vitro cell-free transcription, in vitro transcription of SP6 or T7 RNA polymerase. | |
| Notes | • RNase Inhibitor shows highest activity within pH 7.0-pH 8.0, a wider pH range can inhibit its activity. | |
• Foaming or vigorous vortex can result in the inactivation of RNase Inhibitor. | ||
Instructions for Use
First-strand cDNA synthesis
1. Reaction Components
| Component | Volume |
| Total RNA/mRNA | 50 ng-5 μg/5-500 ng |
| Anchored Oligo(dT)18 Primer (0.5 μg/μl) | 1 μl |
| or Random Primer (0.1 μg/μl) | 1 μl |
| or GSP | 2 pmol |
| 10 mM dNTPs | 1 μl |
| 5× RT Buffer | 4 μl |
| Ribonuclease Inhibitor | 0.5 μl |
| EasyScript®/TransScript® RT | 1 μl |
2. Incubate
| • For anchored oligo(dT)18 primer or GSP, incubate at 42oC for 30 minutes. |
| • For random primer, incubate at 25oC for 10 minutes, then at 42oC for 30 minutes |
| 3. Incubate at 85oC for 5 minutes to inactive EasyScript®/TransScript® RT. |
Packaging
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