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Murine RNase Inhibitor-Avoiding RNase contamination

RNase contamination is always a concern when performing experiments involving RNA. Even with the cleanest of techniques, RNase contamination can happen and may have profound effects on data from downstream applications. High-quality, intact RNA is crucial to the success of sensitive applications. So, are there reagents that prevent RNA from being degraded? Yeah, murine RNase inhibitor of Yeasen is the best choice for solving this problem.

 

1. How to prevent RNA from being degraded?

2. What are RNase inhibitors?

3. What does RNase Inhibitor do?

4. What are the features of Murine RNase Inhibitor?

5. Related products and performance

6. Related products

7. Frequently Asked Questions

 

1. How to prevent RNA from being degraded?

RNase is exceedingly hard to inactivate and exhibits high stability, heat resistance, acid resistance, and alkali resistance. After thermal denaturation, it can quickly regain its original structure. RNase is usually very active and is broadly distributed in both eukaryotic and prokaryotic cells and tissues. The RNA in the sample can completely degrade with just a trace amount of RNase contamination. RNase contamination in studies can come from both internal and external sources. RNase in cells is primarily an endogenous source of contamination, while exogenous sources include experimental supplies, the lab environment, and experimenters themselves.
In order to avoid the degradation of vulnerable RNA, Special precautions must be taken when working with RNA. All reagents and equipment must be specially treated to inactivate RNases prior to use. In addition, RNA inhibitors can be used to inhibit RNase activity and prevent RNase degradation of RNA.

2. What are RNase inhibitors?

RNase inhibitors are a class of substances that can inhibit RNase activity. Common RNase inhibitors include Diethyl Pyrocarbonate (DEPC), Guanidine Isothiocyanate, Ribonucleoside Vanadyl Complexes (RVC), and protein inhibitor of RNase (RNasin). Among them, DEPC and guanidine isothiocyanate has certain toxicity, and the RVC has an inhibitory effect on PCR polymerase, which is not conducive to subsequent experiments. RNasin is not toxic and is a non-competitive inhibitor of RNase, which can bind to various RNases to inactivate them.

3. What does RNase Inhibitor do?

RNase inhibitors are commonly used as a precautionary measure in enzymatic manipulations of RNA to inhibit and control for such contaminants. RNasin is an acidic glycoprotein extracted from rat liver or human blastoderm, which can specifically bind to RNase in a non-covalent manner to form a complex, causing RNase inactivation, thereby protecting the integrity of RNA. At present, the RNase Inhibitor helps to prevent RNA degradation in applications like cDNA synthesis, RT-PCR, in vitro transcription/ translation reactions, or RNA purification.

4. What are the features of Murine RNase Inhibitor?

Recombinant murine RNase inhibitor does not contain 2 oxidation-sensitive cysteine which are contained in human-origin RNase inhibitor. Therefore, murine RNase inhibitor has high anti-oxidation activity and is more stable for low DTT experiments. In addition, Murine RNase Inhibitor of Yeasen has the following features:
1) All-around RNase inhibition: Murine RNase Inhibitor of YEASEN specifically inhibits RNase A, RNase B, RNase C and so on.
2) Versatile reaction conditions: RNase inhibitor is active at pH 5.0-9.0 and 25℃-55℃, which is suitable for thermostable reverse transcriptase.
3) Multiple downstream experiments possible: No inhibition of polymerase activity is observed when RNase Inhibitor is used with Taq DNA Polymerase, AMV or M-MuLV Reverse Transcriptase, or Phage RNA Polymerases (SP6, T7, or T3).
4) Mass-produced goods: a single manufacturing capacity of 2 billion U is helpful to cost management by ensuring product homogeneity, supply stability, and timeliness.
5) Batch-to-batch consistency: mature protein expression and purification platform in accordance with ISO13485 quality management system, quality control testing in accordance with quality requirements to assure product stability between batches.

5. Related products and performance

A. RNase Inhibitor blocks RNase activity

1μg HEK cell total RNA, 1μL RNase inhibitor can effectively inhibit 5ng RNase.

Inhibitory effect of Murine RNase Inhibitor.

Fig 1. Inhibitory effect of Murine RNase Inhibitor.

B. RNase Inhibitor surpasses foreign products in qPCR testing

The inhibitory impact of the Yeasen and R* Company mouse-derived RNase inhibitors (MRI) was measured using the qPCR technique under the identical experimental circumstances, and the RNase inhibitor of the YEASEN Murine source efficiently inhibited RNase A in the system. The impact of inhibition was superior to that of competitors.

Fig 2. R* MRI RNase inhibition test

Note: The curves in the figure from left to right are: Positive control (RNA only, no RNase and MRI), 40 U MRI, 30 U MRI, 20 U MRI, 10 U MRI, Negative control (RNA + RNase, no MRI).

Yeasen MRI RNase inhibition test

Figure 3. Yeasen MRI RNase inhibition test
Note: The curves in the figure from left to right are: Positive control (RNA only, no RNase and MRI), 80 U MRI, 60 U MRI, 40 U MRI, 30 U MRI, 20 U MRI, 10 U MRI, Negative control (RNA+RNase, no MRI).

 The CT of RT-qPCR results of Yeasen MRI and R* MRI

Fig 4. The CT of RT-qPCR results of Yeasen MRI and R* MRI

6. Related products

Product Name

SKU

Specifications

Murine RNase Inhibitor (40 U/µL)

10603ES05

2 KU

10603ES10

10 KU

10603ES20

20 KU

10603ES60

100 KU

10603ES94

20,000 KU

Murine RNase inhibitor (200 U/μL)

10610ES03

1 mL

10610ES50

50 mL

10610ES76

500 mL

Murine RNase Inhibitor (200 U/µL, Glycerol-free)

10703ES05

2 KU

10703ES10

10 KU

10703ES60

100 KU

10703ES70

200 KU

10703ES80

1,000 KU

7. Frequently Asked Questions

Q1: Will there be RNase in Murine RNase inhibitor?
A: Each reagent will detect RNase activity to ensure that Murine RNase inhibitor does not bring RNase contamination.
Q2: Will Murine RNase inhibitor have an impact on downstream PCR experiments?
A: It will have no effect. Each batch of Murine RNase inhibitor is clear of genomic contamination after quality testing and may be used in RT-PCR and RT-qPCR research.
Q3: Will Murine RNase inhibitor be inactivated?
A: Inhibitors will be inactivated at temperatures above 65°C, and vigorous stirring will also cause inactivation.

Regarding reading:

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