DNA purification is a crucial step in many molecular assays which include PCR, qPCR, and DNA sequencing. It removes contaminating proteins as well as salts and other impurities that can hinder the downstream process. It also ensures the desired DNA is completely clean and in good condition so that it can then be used for further analyses. The quality of DNA can be assessed by using spectrophotometry or gel electrophoresis and other methods.
In the initial step of a DNA purification procedure the cellular structure will be disrupted by detergents or reagents such as SDS to release DNA. To further purify the DNA, reagents with protein denature such as sodium dodecylsulfate as well as Ethylene diamine tetraacetic acid (EDTA) are added to denature the proteins. Then, they are removed from the nucleic acid solution using centrifugation and wash steps. If RNA is detected in the sample then it can be further denatured by adding ribonuclease. The nucleic acid is reacted with ethanol that has been cooled to make it distinct from http://www.mpsciences.com/2021/02/15/science-supplies-for-students/ other contaminants.
Ethanol is an everyday solvent that can be used to eliminate salts and other contaminants from nucleic acid samples. Researchers can compare results from different experiments using an ethanol concentration that is standard, which is a good option for workflows that require high-throughput. Other solvents such a chloroform or phenol can be used, but these are more toxic and may require additional steps to avoid cross-contamination with other proteins or cellular debris. Newer methods can make it easier to complete the DNA purification process using low-ionic-strength ethanol, which has been shown to be equally effective as traditional organic solvents in purifying DNA [26]. This is particularly applicable when used in conjunction with spin column extract kits.
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