RNase R |
Catalog No.GE10003
One-Click Copy Product Info
|
RNase R is an E. coli exoribonuclease which exhibits 3’-to-5’ exonuclease activity, efficiently digesting nearly all linear RNA species.
Products are for research use only. Not for human use. We do not sell to patients.
Sample solution is provided at 25 µL, 10mM.
Applications: Circular RNA studies; linear RNA removal from circular RNA; alternative splicing studies; analysis and identification of intron lariat sequences, etc.
Source: E.coli RNase R expressed and purified from E. coli.
Definition of activity: One unit converts 1μg of poly-r(A) into acid-soluble nucleotides in 10 minutes at 37℃ in 20mM Tris-HCl (pH8.0), 100mM KCl and 0.1mM MgCl2.
Purity: Free of DNase, RNA endonuclease and exonuclease.
Enzyme storage buffer: 50mM Tris-HCl (pH7.5 at 25℃), 200mM NaCl, 1mM DTT, 0.1mM EDTA, 50% (v/v) Glycerol, 0.1% (w/v) Triton X-100.
10X RNase R Reaction Buffer: 200mM Tris-HCl (pH8.0 at 25℃), 1M KCl, 1mM MgCl2.
RNase R activity is dependent on magnesium ion at a concentration of 0.1-1.0mM. Chelators such as EDTA will significantly reduce the activity of RNase R. If necessary, add MgCl2 additionally in the reaction mixture to a final concentration of 0.1mM at least.
Instructions for Use:
1. Linear RNA digestion with RNase R
a. Set up the reaction on ice as follows:
|
Reagent |
Volume/Concentration |
|
|
RNA |
<5µg |
>5µg |
|
10X RNase R Reaction Buffer |
2µl |
5µl |
|
RNase R (20U/µl) |
1-3U/µg RNA |
1-3U/µg RNA |
|
DEPC-treated Water |
up to 20µl |
up to 50µl |
b. Mix the reaction mixture well and incubate at 37℃ for 10-30min.
c. Incubate at 70℃ for 10min to terminate the reaction.
Note 1: The amount of RNase R and the volume of the reaction mixture need to be optimized through experiments.
Note 2: The RNase R digestion product after heat-inactivation can be used directly for RT-PCR and qPCR, etc.
2. Purification and recovery of circular RNA by one of the following methods
a. Phenol/chloroform extraction followed by ethanol precipitation.
(a) Add Nuclease-free Water into the reaction mixture obtained at step 1c to a total volume of 180μl. Add 20μl of 3M sodium acetate (pH 5.2) or 20μl of 5M ammonium acetate, mix well. Add 200μl of phenol/chloroform mixture (1:1), vortex vigorously for 20-30s. Centrifugation at 12000rpm for 5-10min and aspirate the upper layer phase into a new centrifuge tube.
(b) Precipitate RNA by adding double volume of absolute ethanol, place at -20℃ for at least 30 minutes, and centrifuge at 12000rpm at 4℃ for 5-10min.
(c) Discard the supernatant and wash the pellet with 500μl of ice-cold 70% ethanol to fully remove the salt.
(d) Dissolve RNA in DEPC-treated Water (R0021/R0022) and store at -80℃.
b. Use RNA purification column or magnetic beads for RNA purification and recovery.
References:
1. Suzuki, H. et al., Nucl Acids Res. 2006; 34(8):e63.
2. Vincent, H.A. and Deutscher, M.P., J Biol Chem. 2006; 281(40):29769.
| Cas No. | SDF | ||
| Formula | M.Wt | ||
| Solubility | Storage | Store at -20℃ | |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Average Rating: 5 (Based on Reviews and 40 reference(s) in Google Scholar.)