Rnase Inhibitor (Glycerol free)
Murine RNase inhibitor is a recombinant murine RNase inhibitor expressed and purified from E.coli. It binds to RNase A, B or C in a 1:1 ratio through non-covalent bonding, thereby inhibiting the activity of the three enzymes and protecting RNA from degradation. However, It is not effective against RNase 1, RNase T1, S1 Nuclease, RNase H or RNase from Aspergillus. Murine RNase inhibitor was tested by RT-PCR, RT-qPCR and IVT mRNA, and was compatible with various commercial Reverse transcriptases, DNA polymerases and RNA polymerases.
Compared to human RNase inhibitors, murine RNase inhibitor does not contain two cysteines that are highly sensitive to oxidation which causes inactivation of the inhibitor. That making it stable at low concentrations of DTT (less than 1 mM). This feature makes it suitable for use in reactions where high concentrations of DTT is adverse to the reaction (e.g. Real-time RT-PCR).
Application
This product can be widely used in any experiment where RNase interference is possible to avoid RNA degradation, such as:
1.First-strand cDNA synthesis, RT-PCR, RT-qPCR, etc.;
2.Protect RNA from degradation in in-vitro transcription/translation (e.g. viral replication in vitro);
3.Inhibition of RNase activity during RNA isolation and purification.
Storage Conditions
Store at -25～-15℃;
Freeze-thaw cycles ≤ 5 times;
Valid for 1 years.
Unit definition
One unit is defined as the amount of RNase Inhibitor required to inhibit the activity of 5 ng of RNase A by 50%.
Molecular weight
RNase Inhibitor (Glycerol-free) is a 50 kDa protein.
Quality control
Exonuclease Activity:
Incubation of 40 U of enzyme with 1 μg λ-Hind III digest DNA for 16 hours at 37°C resulted in no detectable degradation of the DNA as determined by gel electrophoresis.
Endonuclease Activity:
Incubation of 40 U of enzyme with 1 μg λ DNA for 16 hours at 37°C resulted in no detectable degradation of the DNA as determined by gel electrophoresis.
Nicking Activity:
Incubation of 40 U of enzyme with 1 μg pBR322 for 16 hours at 37℃ resulted in no detectable degradation of the DNA as determined by gel electrophoresis.
RNase Activity:
Incubation of 40 U of enzyme with 1.6 μg MS2 RNA for 4 hours at 37℃ resulted in no detectable degradation of the RNA as determined by gel electrophoresis.
E.coli DNA:
40 U of Enzyme is detected by TaqMan qPCR. The E.coli DNA is ≤ 0. 1pg/40U.
Notes
1.Don’t shake or stir violently to prevent enzyme inactivation.
2.RNase inhibitor is active at temperatures ranging from 25℃ to 55℃ and is inactivated at ≥65℃.
3.The activity of RNase H, RNase 1, and RNase T1 is not inhibited.
4.The pH range for inhibiting RNase activity is wide (active at pH 5-9), exhibiting maximum activity at pH 7-8.
