RNaseOFF Ribonuclease Inhibitor
Cat. No.
G138
Unit
4,000 U (100 µl)
Price
$80.00
| Cat. No. | G138 | |
| Name | RNaseOFF Ribonuclease Inhibitor | |
| Unit | 4,000 U (100 µl) | |
| Category | Molecular Biology Enzymes and Kits | |
| Description |
Same performance as competing brands, better pricing!RNaseOFF Ribonuclease Inhibitor specifically inhibits common ribonucleases (RNases), including RNase A, B, and C, with high affinity. This enzyme is a useful additive in PCR or RT-PCR as it safe-guards RNA against potential RNase contamination without inhibiting polymerase activity. This robust version of RNase inhibitor has improved resistance to oxidation compared to the high oxidation-sensitive human RNase inhibitor. RNaseOFF is stable even under very low concentrations of DTT (< 1 mM), making it the best choice for ultimate RNA protection. Product Features:
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| Application |
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| Concentration | 40 U/µl | |
| Storage Condition |
Store at -20°C. |
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| Note |
One unit is defined as the amount of RNaseOFF Ribonuclease Inhibitor that is required to inhibit the activity of 5 ng of RNase A by 50%.
This product is distributed for laboratory research only. Caution: Not for diagnostic use. |
| Are all components in this product RNase-free? | |
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Yes, all components in the RNaseOFF Ribonuclease Inhibitor is RNase-free.
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| What is the shelf life of this reagent? | |
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1 year upon shipment of the reagent.
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| How many reactions can be performed? | |
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The appropriate number of units per reaction will vary depending on the amount of RNase contamination that is present in the reaction mixture for any given experiment.
One unit of RNaseOFF Ribonuclease Inhibitor will inhibit the activity of 5 ng of RNase by 50%; thus, the more RNase contamination present in the specific reaction, the larger number of units of RNaseOFF that will be required for the inhibition.
As a general guideline, a typical amount of RNaseOFF used would be around 25-50 U per 50 µl reaction; note that this is a general guideline only and the number of units required will vary from case to case.
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| Is DTT required for the RNaseOFF Ribonuclease Inhibitor to function? | |
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No. DTT (dithiothreitol) is used to stabilize enzymes and proteins, and it is not required for the RNaseOFF Ribonuclease Inhibitor to function/inhibit ribonucleases.
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| Does RNaseOff (Cat. No. G138) inhibit RNase R activity? | |
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No, RNAseOff does not inhibit RNase R activity.
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| How efficiently will RNase R (when best optimized) digest linear RNA? | |
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To calculate the percentage of digested linear RNA in your experiment, perform the following steps: Step 1. Determine the Ct difference (ΔCt) between the Treated (+RNase R) and Untreated (No RNase R) Set. Step 2. Calculate the fold change using the ΔCt value: Fold Change = 2ΔCt Step 3. Calculate the percentage of RNA degraded using the fold change: % RNA Degraded = (1 - (1 / Fold Change)) × 100
For example, if your Treated Set has Ct=28.55 and Untreated Set has Ct = 18.01, the calculation will be: ΔCt = 28.55 – 18.01 = 10.54 Fold Change = 2ΔCt = 210.54 = 1488.87 % RNA Degraded = (1-(1/1488.87)) × 100 = 99.93% |
| What is the length of RNA that can be digested by RNase R? | |
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RNase R does not have a limit on the length of RNA that it can degrade. Therefore, the majority of non-circular RNA with the exception of short double-stranded RNA, 5S RNA and tRNA will be digested.
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| What inhibits RNase R? | |
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Activity of the enzyme will be abolished if EDTA is present at a concentration of 1 mM or higher. Therefore, if using EDTA compensate by adding MgCl₂ to 1.0 mM final concentration.
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| How is RNase R inactivated/removed? | |
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While the enzyme can be heat inactivated, the procedure is not recommended since high heat can lead to RNA damage. Phenol-chloroform precipitation can be used instead. For NGS, solid phase reversible immobilization (SPRI) bead cleanup is recommended.
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| Can I use higher temperatures (50 degrees) for my sample incubation? | |
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While some of the literature reports that RNase R enzyme is active at temperatures equal or slightly higher than 50°C, we do not recommend exceeding the suggested range of 37-45°C. For NGS applications, incubation at 37°C is recommended. This will ensure optimal activity of the enzyme and absence of non-enzymatic degradation.
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| Can RNase R be used for total RNA isolation in addition to RNA degradation? | |
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Yes, RNase R can be used for both RNA degradation and total RNA isolation. It efficiently degrades linear RNA, leaving circular RNA and genomic DNA intact.
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| Can RNase R be used for RNA integrity assessment in RNA-seq experiments? | |
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Yes, RNase R is commonly used for assessing RNA integrity before RNA-seq experiments. It helps confirm the presence of intact RNA and can be a valuable quality control step.
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| Can RNase R be used for RNA samples in difficult matrices, such as tissues or formalin-fixed paraffin-embedded (FFPE) samples? | |
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RNase R can be used for a variety of sample types, including tissues and FFPE samples. However, sample preparation methods should be optimized for challenging samples (such as incubation time and reactions conditions).
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| Is RNase R suitable for all RNA types, including eukaryotic and prokaryotic RNA? | |
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RNase R is suitable for degrading both eukaryotic and prokaryotic RNA. It is a broad-spectrum ribonuclease.
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| Does RNase R require special precautions for handling and disposal? | |
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RNase R is not hazardous. However, it is important to use appropriate lab safety practices and dispose of waste materials according to local regulations.
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| Is there any risk of contamination of RNase R with DNase or other contaminants? | |
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RNase R is rigorously tested for purity and is free from contaminants, including DNase. It is provided in a ready-to-use format.
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- Del Toro, N., Lessard, F., Tardif, S., Bouchard, J., Bourdeau, V., Ferbeyre, G., & Brakier-Gingras, L. "The retinoblastoma tumor suppressor limits ribosomal readthrough during oncogene induced senescence" bioRxiv 788380: (2019).
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