Production Overview

The RNase detection kit is based on a fluorophore-labeled RNA probe. When the sample does not contain RNase activity, the probe is stable and does not produce a fluorescent signal; when the sample contains RNase activity, the probe is degraded, resulting in a gradual enhanced fluorescence signal; the rate of increase in fluorescence signal is positively correlated with the number and activity of enzymes. Use a fluorescence microplate reader to measure at the wavelength of ex/em=535/575nm to determine whether the sample is contaminated by RNase.

Key Features

• Easy to use, get the result within 30 minutes.

• Test for activity and ensure complete absence of contamination.

Product Highlights

• High sensitivity: Detection limit：RNase A：0.313pg/mL or 1.56×10^-9U/μL

• High accuracy: recovery between 80%-120%

• High precision: intra batch coefficient of variation ≤ 10%, inter batch coefficient of variation ≤ 15%

• High applicability: Benzonase Nuclease，mung bean nuclease, S1 nuclease, RNase T1，RNase I can be detected.

Applications

Assay to detect and quantify RNase in buffer, consumables，purified IVT product, mRNA drug substance.

Performance Data

Compared with one of the best competitor kits (fluorescence), the limit of detection of Hzymes RNase Assay kit is only 1/8 of that of the competitor kit.

Precision and Stability:

Different types of enzymes can all be detected.

Adapt to different instruments.

Custom Case

We have cumulatively facilitated multiple IND applications in both China and the United States, covering cutting-edge fields such as mRNA vaccines, therapeutic mRNA for oncology, and in vivo CAR-T cell therapy.

FAQ

When could false positive or false negative results or inaccurate quantitative results occur?

a. Gel buffers, high concentration of viscous substances, surfactants and dark solutions may interfere with the luminescence of fluorophores;

b. If the sample solution to be tested contains substances that inhibit RNase activity, the result of the measurement is the overall enzyme activity of the sample solution, not the activity of the enzymes within it. These substances include:

• High ionic strength solutions (eg.5M NaCl,20x SSC,3M sodium acetate, etc.)

• Buffers with pH<4 or pH>9

• Chaotropic agents, detergents, chelating agents or any solution that denatures proteins (eg. SDS, guanidine thiocyanate, urea, EDTA,etc. )

c. Solutions that cause chemical instability of RNA probes, such as pH>9 solutions, caustic solutions (strong acids and bases, bleach, etc.)

While preparing standard solutions, in the pre-treatment steps why use the standard dilution buffer instead of DNase &RNase-free water to dilute?

Because the standard is relatively stable in the standard dilution buffer even after series dilution. But if diluted with water, the activity of the standard may change during the series dilution process, resulting deviation of the standard.

Why seriously contaminated samples may lead to false negative results?

The criterion for judging RNase contamination is: RFU30 (sample to be tested)≥2×RFU0 (sample to be tested), that is, the RFU value at 30 minutes of reaction is more than twice the RFU value at 0 minute. If the sample is seriously contaminated, it is possible that the reaction starts very quickly, so a very high RFU0 value is measured in a very short time, resulting in a false negative result of RFU30 (sample to be tested)<2×RFU0 (sample to be tested). At this time, the sample to be tested needs to be diluted with DNase & RNase-free water.

If the RFU value of the negative control is not 0, does it mean that the negative control is contaminated?

Uncertain. The negative control can also be detected with a low RFU signal (base line value) but the value will not increase significantly along with the proceeding of the reaction. Generally, it is considered that the RFU30 is less than twice the RFUO which indicates that the negative control is not contaminated.