Precautions for RNA Extraction Using a Low-Temperature Centrifuge

RNases are ubiquitous, and at any stage of experimental procedures, even the slightest oversight or improper handling can lead to RNase contamination, thereby jeopardizing the entire experiment. Therefore, strict control of experimental conditions and avoidance of any potential contamination are crucial for ensuring the success of an experiment. To this end, the following measures must be taken:

1. If possible, the laboratory should designate a separate area specifically for RNA handling. All equipment—including Lu Xiangyi low-temperature centrifuges, pipettes, and reagents—should be used exclusively in this RNA handling area. The RNA handling area must be kept clean and regularly sterilized.

2. If possible, perform the procedure in a clean bench according to cell culture requirements to effectively avoid RNAase contamination caused by handling.

3. During the procedure, always wear disposable rubber gloves and change them frequently to prevent bacteria and fungi from your hands and arms—as well as RNA enzymes naturally secreted by your body—from being transferred to test tubes or contaminating laboratory equipment. Avoid using disposable plastic gloves whenever possible. Plastic gloves not only often cause operational inconvenience but also tend to transfer RNase from contaminated areas of equipment to RNase-free areas, thereby exacerbating contamination.

4. Avoid talking or chatting during the procedure. You can also wear a mask to prevent RNAase contamination.

5. As much as possible, use single-use plastic products and avoid sharing equipment such as filter papers, tips, and tubes to prevent cross-contamination. For example, researchers working with RNA probes often use enzymes like RNase H and T1; during the experimental process, there is a high likelihood of contaminating pipettes, centrifuges, and other equipment. Such contaminated equipment poses a significant threat to RNA manipulation.

6. Regarding single-use plastic products, it is recommended to use tips and tubes supplied by manufacturers that have already been sterilized prior to shipment. Most manufacturers’ sterile plastic products rarely contain RNAase contamination; once purchased, they can be used directly for RNA manipulation. Many researchers who use plastic products treated with DEPC or similar agents often find that these products become contaminated with RNAase due to secondary contamination, thereby leading to experimental failures.

7. Alcohol, isopropyl alcohol, Tris, and other reagents used for preparing solutions should be from unopened, new bottles.

8. All old plastic products must be treated with 0.5 M NaOH for 10 minutes, thoroughly rinsed with double-distilled water, and then sterilized after being immersed overnight in DEPC-treated H2O.

9. Instruments that cannot be treated with DEPC can be wiped several times with chloroform; this usually effectively inactivates RNases. However, chloroform can dissolve certain plastic materials, so caution is advised.

10. RNase AwayTM reagent can replace DEPC; it is easy to use, inexpensive, and non-toxic. Simply pour RNase AwayTM directly onto the surfaces of glass and plasticware, let it soak for a few minutes, then rinse thoroughly with water to remove it. This quickly eliminates RNase from the surface of your labware without leaving any residue that could interfere with subsequent experiments.

11. If you need to transport RNA samples over long distances or store them for extended periods, we recommend first preserving the specimens (cells or tissues) in an RNA preservation solution (such as RNAwait or RNAlater). This allows the RNA within the cells to be separated from RNA enzymes, enabling storage at room temperature for up to 7 days, at 4°C for up to 4 weeks, and at -20°C or -80°C for long-term archival storage without compromising RNA quality. Even after such long-term storage, high-quality RNA can still be efficiently extracted using various methods.