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7/13/2021 | posted: 6/13/2022 at 9:07:42 AM ET
Achieving complete isolation of nucleic acids is not always easy. The methods used to isolate these molecules vary depending on the type of samples and their concentration. Animal, plant, and microbial samples can all present a number of obstacles that must be overcome to achieve complete purification. These barriers include the presence of inhibitors, contaminants, and trace amounts of the target organism. In these cases, it is necessary to process the samples with different steps.
Typically, the Nucleic Acid purification with the use of magnetic carriers with affinity ligands. These particles are made of biopolymers or synthetic polymers, porous glass, and inorganic magnetic materials. In addition to their magnetic properties, these particles can also be modified using ligands that can selectively capture nucleic acids. The process can be used to separate RNA and DNA, or to isolate nucleic acids from other molecules such as proteins, carbohydrates, or lipids.
The magnetic separation technique has many advantages over other methods of nucleic acid purification. One of the advantages of this technique is that the nucleic acids can be isolated directly from the crude sample materials, such as blood, tissue homogenates, and cultivation media. It also allows for the recovery of small particles that are otherwise difficult to extract. The method is also highly efficient, fast, and automated, allowing the purification of nucleic acids from even the largest samples.
To separate nucleic acids from the matrix of polyacrylamide gels, different methods are used. One method involves phenol extraction followed by ethanol precipitation. However, phenol is toxic, so it is disposed of as organic waste. The other popular method uses silica-based columns. A special agarose gel is used to separate specific bands. Samples are loaded into the top row of the wells, while the bands are retrieved from the bottom row.
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