how to remove phenol contamination in rna

How to remove RNA in phenol pollution: detailed analysis and methods

In the process of RNA extraction, phenol is a common organic solvent, which can effectively remove lipids and proteins from cells. The residue of phenol may affect the results of subsequent experiments, especially the purity and stability of RNA. Therefore, how to remove phenol contamination in RNA is an important issue in molecular biology experiments. This article will analyze in detail how to effectively remove phenol contamination in RNA and provide a feasible method.

Phenol Contamination on RNA Experiment

Phenol contamination can affect the quality and purity of RNA. Phenol is used to lyse cells and isolate RNA during RNA extraction, but if not handled properly, phenol may remain in the RNA sample. Phenol residues not only interfere with the absorbance determination of RNA, but also may lead to RNA degradation, thus affecting the follow-up experiments, such as reverse transcription PCR(RT-PCR) and Northern Blot. Phenol is strongly irritating and corrosive, and residual phenol also poses a certain risk to the health of laboratory personnel.

How to remove RNA from phenol contamination

Removal of phenol contamination in RNA generally relies on several common methods:

1. Centrifugation layered extraction

Centrifugal stratified extraction is the most common method to remove phenol contamination. During RNA extraction, after the aqueous phase is separated from the organic phase, further separation can be carried out by centrifugation. RNA is usually present in the aqueous phase, while phenol and other organic solvents accumulate in the organic phase layer. In operation, an appropriate centrifugation speed and time can be used to ensure complete separation of the phenol layer from the RNA layer. This method is simple and effective, and can effectively remove phenol pollution.

2. Add chloroform for secondary extraction

After RNA extraction, chloroform may be added for a second extraction. Chloroform is mixed with phenol to further remove phenol and other impurities from the sample. In operation, after adding chloroform, the sample was gently shaken and centrifuged to separate the aqueous, phenol and chloroform layers. This method can remove phenol contamination more thoroughly and reduce the loss of RNA purity.

3. Dialysis to remove phenol

If the phenol contamination in the RNA sample is serious, it can be removed by dialysis. Through the selective permeability of the dialysis membrane, the phenol molecules in the solution can be effectively removed. The advantage of the dialysis method is that it can remove phenol contamination while ensuring that RNA is not damaged, and it is especially suitable for samples with high phenol residue.

Other phenol removal methods

In addition to regular of the above methods, there are some other techniques for removing phenol, such:

• Resin adsorption method: through the use of specialized resin to adsorb phenol, can not affect the RNA of the premise to remove phenol pollution.
• Ultrafiltration: the use of appropriate molecular weight cut-off ultrafiltration membrane, can effectively remove phenol and other small molecular pollutants.

These methods can be selected according to specific experimental needs.

Conclusion: how to remove RNA in phenol pollution?

Removal of phenol contamination in RNA is a critical step to ensure RNA purity and experimental success. regular methods, such as centrifugal extraction, adding chloroform for secondary extraction, dialysis, combined with the specific conditions of the laboratory, can effectively remove the phenol pollution in RNA, so as to improve the reliability and accuracy of the experimental results. Mastering these methods of removing phenol contamination is essential to ensure the high quality of molecular biology experiments.