how to remove phenol contamination in rna

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

In the process of RNA extraction, phenol is a common organic solvent-based products, which is able to efficiently remove lipids and proteins from cells. The residue of phenol might affect the results of subsequent experiments, especially the purity and stability of RNA. Therefore, how to remove phenol contamination in RNA is an crucial issue in molecular biology experiments. This article will examine in detail how to efficiently remove phenol contamination in RNA and provide a feasible method. But Effect of Phenol Contamination on RNA Experiment

Phenol contamination is able to affect the condition and purity of RNA. Phenol is applied to lyse cells and isolate RNA during RNA extraction, however if not handled appropriately, phenol might remain in the RNA sample. Makes sense, right?. In particular Phenol residues not only interfere with the absorbance determination of RNA, however also might lead to RNA degradation, thus affecting the follow-up experiments, such as reverse transcription PCR(RT-PCR) and Northern Blot. Moreover 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. Generally speaking 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 is able to be carried out by centrifugation. RNA is usually present in the aqueous phase, while phenol and other organic solvents build up in the organic phase layer. In operation, an appropriate centrifugation speed and time is able to be applied to ensure complete separation of the phenol layer from the RNA layer. In my experience, This method is simple and efficiently, and is able to efficiently remove phenol contamination. And From what I've seen,

2. But Add chloroform to secondary extraction

After RNA extraction, chloroform might be added to 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. Crazy, isn't it?. This method is able to remove phenol contamination greater thoroughly and minimize the loss of RNA purity. And In my experience,

3. Dialysis to remove phenol

If the phenol contamination in the RNA sample is serious, it's able to be removed by dialysis. For example Through the selective permeability of the dialysis membrane, the phenol molecules in the solution is able to be efficiently removed. The advantage of the dialysis method is that it's able to remove phenol contamination while ensuring that RNA isn't damaged, and it's especially suitable to samples with high phenol residue. Additionally Other phenol removal methods

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

Resin adsorptive processes method: through the consumption of specialized resin to adsorb phenol, is able to not affect the RNA of the premise to remove phenol contamination. And Ultrafiltration: the consumption of appropriate molecular weight cut-off ultrafiltration membrane, is able to efficiently remove phenol and other small molecular contaminants. These methods is able to be selected according to specific experimental needs. You know what I mean?. summary: how to remove RNA in phenol contamination?

Removal of phenol contamination in RNA is a critical measure to ensure RNA purity and experimental success. And First regular methods, such as centrifugal extraction, adding chloroform to secondary extraction, dialysis, combined with the specific conditions of the laboratory, is able to efficiently remove the phenol contamination in RNA, so as to enhance the reliability and accuracy of the experimental results. But Mastering these methods of removing phenol contamination is essential to ensure the high condition of molecular biology experiments.