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A Complete Guide To DNA

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Deoxyribonucleic acid (DNA) is basically a hereditary material in human beings and many other organisms. Almost all cells in a person’s body have the same deoxyribonucleic acid.

Most deoxyribonucleic acid can be found in the cell nucleus, and a small amount of it is located in the mitochondria, giving it the name mtDNA (mitochondrial DNA).

Usually, information in the DNA is stored as a code consisting of four bases. These include thymine, cytosine, guanine, and adenine.

The human DNA comprises around 3 billion bases, and about 99% of these bases are similar in every individual.

The sequence or order of those bases helps to determine the information available for maintaining and building organisms, similar to how alphabetical letters appear in a particular order to form sentences and words.

What Deoxyribonucleic Acid Looks Like

According to the history of DNA, James Watson and Francis Crick published a certain article showing the shape of deoxyribonucleic acid as a double helix.

These two were not able to see the deoxyribonucleic acid directly since it is too minuscule for that. But they made a conclusion based on X-ray diffraction images and calculations.

The Basics of DNA Purification

There are several basic steps of extracting DNA, which are consistent across every possible purification chemistry. Some of these include:

Creating Lysate

This involves releasing deoxyribonucleic acid into a solution. The key goal of this step is to completely and rapidly disrupt cells in a sample so as to release DNA into a lysate. There are general techniques for lysing materials, including:

  • Chemical methods
  • Enzymatic methods
  • Physical methods

Clearing Lysate

Based on the starting materials, the cellular lysate might need to have the debris removed before the purification of DNA so as to minimize the carryover of the unwanted materials, such as lipids and proteins. Normally, clearing is achieved by bead-based filtration or centrifugation methods.

Binding

No matter the technique employed to create cleared lysates, the DNA might get isolated using different techniques. Some experts provide genomic systems for isolating DNA depending on sample lysis.

All these chemistries may influence the purity and efficiency of isolation, and each has a binding capacity property.

Washing

In general, a wash buffer has alcohol, and you can use it to remove salts, proteins, and other contaminants in a sample containing DNA. In addition, alcohol may associate DNA with the matrix.

Elution

Basically, DNA is soluble in a low-iconic-strength solution, like nuclease-free water or TE buffer. When an aqueous buffer is applied, the deoxyribonucleic acid gets released from silica, allowing you to collect the eluate.

Advances in the Sequencing Technologies

When preparing the human genome, scientists used to employ a method involving the sequencing of short overlapping DNA fragments that covers the whole chromosome.

The fragments were then aligned based on the overlapping sequencing, allowing scientists to reconstruct the sequence for every chromosome.

Fortunately, advancements in sequencing technologies make things possible. The current technologies can now sequence DNA fragments that range from several kilobase pairs to more than 100 kilobase pairs.

Final Say!

DNA is a molecule that carries genetic information on organisms, including human beings. Today, the ability to sequence deoxyribonucleic acid at a low cost and high throughput enables the development of several sequencing-based applications and techniques.

 

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