Are nucleic acid experiments unstable? It may be that enzymes are causing trouble.

01 Characteristics and origins of DNase and RNase

DNase and RNase are hydrolases that specifically degrade DNA and RNA, respectively. They break the nucleic acid chain by cutting the phosphodiester bonds in the nucleic acid molecule, leading to degradation. RNase has extremely high stability due to its disulfide bond structure, can resist high temperature boiling and even denaturants, and can refold and restore activity after inactivation, making it the number one natural enemy of RNA-related experiments. Sources of pollution are mainly divided into two categories:

✅ Experimenters: Skin, sweat, saliva (droplets) and hair contain large amounts of nucleases.
✅ Experimental equipment: pipettes, pipette tips, centrifuge tubes, glassware, experimental tables and instrument buttons can easily attach nuclease if not handled properly.
✅ Reagents and water: Ordinary distilled or deionized water and chemical reagents may be contaminated during the production and storage process.
✅ Dust particles in the air: can carry nuclease and fall into open reagents or samples.

✅ Endogenous contamination refers to nucleases carried by the sample itself. All biological tissue and cell samples contain endogenous nucleases. Once the sample leaves the living body or its original growth environment, endogenous enzymes will begin to degrade RNA/DNA. The rate of degradation depends on the endogenous enzyme content and the ambient temperature. For example, liver, pancreas, spleen, thymus, brain tissue, and plant tissue are rich in endogenous nucleases. If not processed properly, nucleic acids will be rapidly degraded.

In view of these potential risks, choosing reliable enzyme-free consumables is the first step to prevent and control pollution. All products of Aijin Biotech, from centrifuge tubes, tips, deep well plates to PCR plates, cell culture series, etc., are manufactured in a 100,000-level dust-free workshop. They are strictly tested before leaving the factory to ensure no DNase/RNase contamination, no pyrogens, and no PCR inhibitors. This allows scientific researchers to focus on the experiment itself without worrying about external risks.

02 Establish a DNase-free and RNase-free experimental environment

Whenever possible, set up a dedicated clean bench or area for RNA/DNA operations. Regularly wipe down countertops and instrument surfaces with nuclease remover. These scavengers can effectively inactivate nucleases and are usually non-toxic and non-corrosive. After use, they need to be wiped with enzyme-free water to remove residues.

Glass and metal vessels can be sterilized by dry baking or treated with nuclease scavengers, which is more convenient and safer.

Prioritize the purchase of commercial, DNase/RNase-free consumables, which are more reliable and safer to use right out of the package.

All consumable products of Aijin Biotech have passed the ISO13485 quality management system and CE certification. The production base covers an area of ​​26,000 square meters and is equipped with 5,500 square meters of 100,000-level and 500 square meters of 10,000-level dust-free workshops. Production and quality control are strictly in accordance with standard processes, which can provide laboratory personnel with truly 'ready-to-use' enzyme-free consumables.

Wear clean powder-free gloves and a mask before experimenting. Change gloves immediately after contact with any potential source of contamination (such as doorknobs, phones, your own skin or hair) and try to avoid talking when handling RNA samples.

It is best to use filter tips in all steps involving reagents. Its high-efficiency filtration and strong hydrophobicity can effectively prevent aerosols from contaminating the inside of the pipette and avoid cross-contamination.

Whenever possible contamination is suspected, gloves, centrifuge tubes, and pipette tips should be changed immediately. Pack the reagents into small portions and use separate bottles for each experiment to avoid contamination of the entire bottle of reagents due to repeated use.

For endogenous contamination, the key is to quickly and effectively inhibit nuclease activity in the sample.

'Quick processing and proper storage of samples'

Samples should be processed as soon as possible after collection. If the nucleic acid cannot be extracted immediately, rapid freezing measures (such as direct injection into liquid nitrogen) must be taken to minimize the degradation of RNA/DNA. For tissue samples, liquid nitrogen milling is the most effective method to disrupt the tissue and simultaneously inhibit endogenous enzymes.

Effectively protect sample RNA from degradation by endogenous enzymes, such as RNAlater, etc.

During the nucleic acid extraction stage, a lysis buffer that can quickly inactivate endogenous nucleases should be used. For tissues with high endogenous enzyme content (such as liver, pancreas, plant tissue, etc.), it is recommended to use phenol-containing lysis buffer to enhance inactivation ability. The electric homogenizer is used in conjunction with liquid nitrogen grinding to ensure that tissue cells are homogenized quickly and thoroughly, so that the active ingredients in the lysis solution immediately inhibit nucleases in the cells.

Aijin Biotech provides centrifuge tubes and liquid storage consumables in a variety of specifications, providing matching solutions for different sample types to prevent contamination throughout the entire process from sample processing to nucleic acid extraction.

It is recommended to use DNase/RNase-free reagents and ultrapure water.

For self-prepared reagents, be sure to use enzyme-free water and treated vessels.

All Aijin Biotech's consumables are free of DNase/RNase contamination, pyrogen-free, and PCR inhibitor-free, and can be safely used in highly sensitive nucleic acid experiments.

03 DNase/RNase-free environment inspection

It is also critical to regularly check the DNase/RNase-free environment in the laboratory.

Incubate a complete RNA or DNA standard fragment with the water or buffer to be verified, and observe it through agarose gel electrophoresis after a period of time.

A clear band indicates the absence of contamination, while tailing or disappearance of the band indicates degradation.

Using commercially available high-sensitivity fluorescent substrate detection kits, extremely small amounts of RNase or DNase activity can be quickly and quantitatively detected, helping to accurately locate the source of contamination.

04 Emergency treatment for nuclease contamination

Once nuclease contamination is suspected or confirmed:

Discard laboratory consumables such as pipette tips and centrifuge tubes that may be contaminated, and use a new batch of reagents.

Use special nucleic acid remover to wipe the table and instruments, and irradiate the experimental area with ultraviolet light.

Aijin Bio's consumable product series covers more than 900 types, including deep well plates, pipette tips, columns, liquid storage tanks, PCR series, cell culture series, etc., which fully meet the enzyme-free prevention and control needs of different experimental scenarios and build a long-lasting and stable clean system for the laboratory.

05 Choose Aijin, choose quality

Maintaining a DNase/RNase-free environment is the basis for ensuring reliable molecular biology experimental results. This not only relies on the standard awareness of the experimenters, but also needs to be guaranteed by reliable experimental consumables.