Background

With the rapid rise of mRNA vaccines and nucleic acid therapeutics, maintaining nuclease-free environments has become critical. Deoxyribonucleases (DNases) and ribonucleases (RNases) are widely present in laboratory environments, consumables, and biological materials. Even trace amounts of these enzymes can degrade RNA or DNA, jeopardizing experimental results and production quality.

The Hzymes RNase Assay Kit offers a high-sensitivity, fluorescence-based solution for detecting RNase or DNase contamination, helping researchers and manufacturers maintain stringent quality control standards in mRNA synthesis, vaccine production, and molecular diagnostics.

Detection Method

The method of DNase and RNase Assay kits are the same, Fluorophore-labelled DN/probes or RNA probes are designed and mixed with the test sample.

When the sample does not contain DNase or RNase activity, the probe is stable, the distance between the fluorophore and the quencher group is relatively small, no fluorescent signal is generated due to the principle of fluorescence resonance energy transfer, When the sample contains DNases or RNases activity, the probe is degraded, the fluorophore and the quencher group move away from each other, resulting in a gradually enhanced fluorescent signal:

The rate of increase in fluorescence signal is positively correlated with the number and activity of the enzyme

Detection method is shown as following:

Fig. 1 Schematic diagram of the principle of the fluorescent probe method

Figure 2. Schematic diagram of the DNase/RNase detection kit reaction curve

Product Pictures

RNASE ASSAY KIT (FLUORESCENCE)

It is used to detect RNase contamination in the environment and materials in scientific research and production processes.

Application

It is used to detect DNase contamination in the environment and materials in scientific research and production processes.

Characteristics of Products

l  High sensitivity: the detection limit for RNase A is as low as 0.313pg/mL or about 1.56x10^-9U/uL which is only 1/8 of the best competitor kit.

l  High precision: intra-batch CVS≤10%, inter-batch CVS≤15%.

l  Good stability: valid for one year at -20℃. After the RNA probe solution is prepared, aliquot and store at -20°C.

Precision and Stability of Hyzmes RNase Assay Kit

Sensitivity higher than gel electrophoresis method

The Hzymes RNase Assay kit was in comparison with the nucleic acid hydrolysis-gel electrophoresis method, the same low-concentration enzyme samples were measured as shown in the figure below.

Hzymes RNase Assay kit measures 2.5x10^-8U/uL RNase A, which can be clearly distinguished from 0 concentration.

Determination of RNase A by electrophoresis. No significant difference between 1st ~8th channels (5x10^-8U/uL) and 17th18th channels (2.5x10^-8U/uL), 19th channel (negative control).

Sensitivity: Hzymes RNase Assay kit VS competitor kit

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.

When the sample concentration is 3.13x10^-9U/uL (low-value sample) the signal intensity of the competitor's RNase detection kit is indistinguishable from the negative control.

When the sample concentration is 3.13x10^-9U/uL(low-value sample)the signal intensity of the Hzymes’ RNase Assay Kit is clearly distinguishable from the negative control.

When the sample concentration of RNase A is from 1x10^-6 to 10x10^-6pg/mL, the Signal-to-voise (S/N) values of the competitor's DNase detection kit is between l to 2, however the signal intensity of Hzymes’ DNase Assay Kit is positive correlated to the concentration of RNase l.

Composition of the Hzymes DNase Assay Kit

Note: The standard is RNase A. The unit for RNase A activity is defined as the amount of enzyme that increases 1.0 in the absorbance at 260 nm of yeast RNA at pH 5.0 and 37 'c [1], 50 RNase A activity units are equivalent to l Kunitz unit [2].

Ordering

References

[1] Thermo scientific RNase A (DNase and Protease-free) User Guide

[2] Kunitz, M. A. A spectrophotometric method for the measurement of ribonuclease activity[J]. journal of Biological Chemistry,1946.3(2):308-320.