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Description
RiboSafe RNase Inhibitor is a recombinant protein which completely inhibits a broad spectrum of eukaryotic RNases, including RNases A, B and C.
Product Highlights
Efficient – effectively inhibits a broad spectrum of eukaryotic RNases, including RNases A, B and C
High-quality - certified free of contaminating DNases, RNases, phosphatases and nickases for improved sample security
Robust – active up to 55 oC across a wide range of DTT concentrations and pH
Flexible – compatible with all common reverse transcriptases, bacterial polymerases and thermostable polymerases
Product Description
Ribonucleases (RNases) are ubiquitous and can be introduced into experiments in many ways: for example co-purification during RNA isolation, carryover from bare hands and pipette tips. This RNase contamination can often go unnoticed. RiboSafe RNase Inhibitor is ideal for RNA-sensitive applications such as RT-qPCR as even a small amount of RNase can be detrimental to the final experimental outcome.
RiboSafe RNase Inhibitor is a highly efficient inhibitor of a broad spectrum of eukaryotic RNases and shows no inhibition of polymerase or reverse transcriptase activity, so can be used in cDNA synthesis or one-step RT-qPCR reactions.
RiboSafe RNase Inhibitor is tested for, purity (run on SDS-PAGE gel), presence and absence of endonucleases, nickases and exonucleases, thermostability, inhibition at different pH levels and activity in the presence or absence of DTT. This quality control allows RiboSafe RNase Inhibitor to be used in highly-sensitive techniques such as single-cell RT-PCR, in vitro RNA synthesis and in vitro translation.
Applications
RNA purification
cDNA synthesis
One-step RT-PCR
One-step RT-qPCR
In vitro transcription/translation
RNA Sequencing
RNase protection assays
Enzymatic RNA labeling reaction
Superior RNA protection
Either RNase A and RNase inhibitor were incubated directly with yeast total RNA (blue column) or RNase A and RNase inhibitor were mixed and pre-incubated before addition to yeast total RNA (grey column). In both cases using RiboSafe RNase Inhibitor blocks more of the RNase A activity than RNase inhibitors from Supplier S.
High efficiency
HeLa cell total RNA was incubated at 37°C for 30 min with RiboSafe RNase Inhibitor in the presence of between 2 ng - 125 pg of RNase A (Lanes 1-4), no RiboSafe RNase Inhibitor and 125 pg of RNase A (Lane 5) and no RiboSafe RNase Inhibitor, no RNase A (Lane 6). Even at high concentrations of RNase A, RiboSafe RNase Inhibitor showed no visible levels of degradation.
No detectable effect on polymerase activity
A two-fold serial dilution of total HeLa cell total RNA (1 µg - 0.075 µg) was reverse transcribed in the presence (Lanes 1-5) and in the absence (Lanes 7-11) of RiboSafe RNase Inhibitor, followed by the amplification of a 1 kb fragment of the Angiotensin receptor II gene using MyFi Mix. There was no detectable difference in PCR product in the presence or absence of RiboSafe RNase Inhibitor.
Effect of temperature on RiboSafe RNase Inhibitor activity
2 µg aliquots of mouse total RNA was incubated at 37oC with RiboSafe RNase Inhibitor and no RNase A (lane 1), no RiboSafe RNase Inhibitor and RNase A (lane 2), or with RiboSafe RNase Inhibitor and RNase A at 37oC, 42oC, 45oC, 50oC and 55oC (lanes 3-7 respectively). Regardless of the temperature, there is no visible levels of degradation when incubated with RiboSafe RNase Inhibitor.
All kit components should be stored at -20°C upon receipt. When stored under the recommended conditions and handled correctly, full activity of the kit is retained until the expiry date on the outer box label.
