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General Steps of DNA Extraction

The document outlines the basics of DNA extraction, a procedure to isolate DNA from cells, detailing five general steps: breaking cells open, separating DNA from proteins, precipitating DNA with alcohol, cleaning the DNA, and confirming its presence and quality. It also mentions that while ready-to-use DNA extraction kits are available, many labs prefer their own cost-effective methods. Extracted DNA can be utilized for various molecular analyses such as PCR, electrophoresis, sequencing, fingerprinting, and cloning.

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3 views2 pages

General Steps of DNA Extraction

The document outlines the basics of DNA extraction, a procedure to isolate DNA from cells, detailing five general steps: breaking cells open, separating DNA from proteins, precipitating DNA with alcohol, cleaning the DNA, and confirming its presence and quality. It also mentions that while ready-to-use DNA extraction kits are available, many labs prefer their own cost-effective methods. Extracted DNA can be utilized for various molecular analyses such as PCR, electrophoresis, sequencing, fingerprinting, and cloning.

Uploaded by

aminqasm111
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd

Molecular Biology Dr. Salam A. Ahmed practical lab.

Basics of DNA Extraction

DNA extraction is a routine procedure used to isolate DNA from the nucleus of cells.

Scientists can buy ready-to-use DNA extraction kits. These kits help extract DNA from
particular cell types or sample types. However, they can be expensive to use routinely, so many labs
have their own methods for DNA extraction.

General Steps of DNA Extraction

Step 1. Breaking cells open to release the DNA

The cells in a sample are separated from each other, often by a physical means such as grinding
or vortexing, and put into a solution containing salt. The positively charged sodium ions in the salt
help protect the negatively charged phosphate groups that run along the backbone of the DNA.

A detergent is then added. The detergent breaks down the lipids in


the cell membrane and nuclei. DNA is released as these membranes are disrupted.

Step 2. Separating DNA from proteins and other cellular debris

To get a clean sample of DNA, it’s necessary to remove as much of the cellular debris as
possible. This can be done by a variety of methods. Often a protease (protein enzyme) is added to
degrade DNA-associated proteins and other cellular proteins. Alternatively, some of the cellular debris
can be removed by filtering the sample.

Step 3. Precipitating the DNA with an alcohol

Finally, ice-cold alcohol (either ethanol or isopropanol) is carefully added to the DNA sample.
DNA is soluble in water but insoluble in the presence of salt and alcohol. By gently stirring the alcohol
layer with a sterile pipette, a precipitate becomes visible and can be spooled out. If there is lots of
DNA, you may see a stringy, white precipitate.

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Molecular Biology Dr. Salam A. Ahmed practical lab.3

Step 4. Cleaning the DNA

The DNA sample can now be further purified (cleaned). It is then resuspended in a
slightly alkaline buffer and ready to use.

Step 5. Confirming the presence and quality of the DNA

For further lab work, it is important to know the concentration and quality of the DNA.

Optical density readings taken by a spectrophotometer can be used to determine the


concentration and purity of DNA in a sample. Alternatively, gel electrophoresis can be used to show
the presence of DNA in your sample and give an indication of its quality.

What can this DNA be used for?

Once extracted, DNA can be used for molecular analyses including PCR, electrophoresis,
sequencing, fingerprinting and cloning.

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