DNA extraction is one of the most important processes involved in the lab. And with good reason. Isolating DNA samples from different biological surfaces is essential to study various organisms and determine the cause of diseases that are harming the human body. This helps in prompt diagnosis and development of drugs that can combat harmful health conditions.
DNA extraction is usually done by disrupting the cell wall or membrane of a particular organism’s cellular components ro collect DNA material from them. The methods used can be chemical, physical, or enzymatic. DNA can be collected from a number of sources such as:
- Body fluids
- Frozen tissue sections
- Buccal swab
How Does DNA Extraction Work?
No matter what the process is used to extract DNA, the actual process remains more or less the same. Let us take a look at the basics of DNA extraction.
Lysis Of The Cell
Through a number of physical disruption methods, such as sonication, freezing and grinding, and vertexing, DNA is released into the lysate. It is then added to a salt solution so that the negatively charged phosphate beads are not destroyed or denatured. In addition, lipid breakdown is often facilitated by adding sodium dodecyl sulphate to the mixture to achieve good results.
The reagents are pipetted using different instruments and high-quality Beckman filter tips to prevent contamination and achieve good results.
Removal Of Cellular Debris
When you extract DNA from a particular organism, it also has other materials and cellular debris in it. This could be a number of unwanted products, such as DNA-associated proteins or cellular proteins. It is no secret that if these particles are not removed, the quality of your experiment will be compromised and the results will not be as accurate.
Cellular debris can be removed through a variety of methods, such as centrifugation, bead-based methods, or adding a relevant protease to the solution containing the sample. Any method depending on accuracy and ease can be chosen. Lab techs should carefully weigh the pros and cons of each.
Precipitation Of DNA
Once the extra debris and material is removed, you can start precipitation of the DNA. This involves centrifuging the solution after adding ice-cold ethanol or or isopropyl alcohol.
The mix of alcohol and salt causes the DNA to become insoluble, hence ammonium or sodium acetate are also added for effective precipitation of nucleic acids and to achieve better results.
Purification Of DNA
Once the DNA pellet is extracted, it is cleaned thoroughly using cold alcohol and then centrifuged. This way, all contaminants are removed from it. Now, the DNA sample can be used to perform different experiments.
While using eluting buffer solutions, it is important to choose the correct one. We would recommend using one that meets the requirements of your downstream application. For example, EDTA may negatively impact a lot of DNA samples as it binds to metal ions and may denature or inhibit the activity of enzymes present in the DNA.
Confirmation Of DNA
Now that you are done with the whole process, it is important to check once whether the DNA is present or not. Electrophoresis is an important process that helps check the quality and presence of DNA.
DNA Electrophoresis is done on an agarose gel and is checked through UV light. In addition, the concentrations can also be checked through this method using a spectrophotometer.
To Wrap It Up
DNA extraction is an important process and has multiple applications, such as in newborn screening, medical diagnosis, genetic engineering, and drug research. Hence, it is important that this process be performed with extreme care and accuracy to avoid any mistakes and discover better things for mankind.
An important aspect of DNA extraction is the use of high-quality equipment. This is exactly where Molecular Biology Products come in. Whether you need filter tips for biomek, well plates, or conical flasks to carry out DNA extraction and numerous other procedures, we have all this and more to suit your needs. Our products are high in quality and will help you yield outstanding results.