Sequencing Sanger is considered to be a very important method that helps to find information about the order and identify the four nucleotide bases in a specific DNA segment. This particular method is also known as the chain termination method.
In the year 1977, this method was developed by Frederick Sanger along with his colleagues. Till now, this is regarded as the gold standard of sequencing technology.
Some of the most important benefits of this technology are flexibility, long read capabilities along a high degree of accuracy. As a result, the sequencing Sanger methods help to effectively support a wide range of applications in many of the research areas.
This particular method was used for sequencing the entire human genome.
Overview Of Sequencing Sanger
- Sanger sequencing generally targets a particular region of the template DNA by making use of an oligonucleotide sequencing primer. As a result, this effectively binds to the DNA that is adjacent to the particular region of interest.
Also, there must be the presence of an area of known sequence that is present close to the target DNA.
- For finding out the sequence, this sequencing Sanger mainly uses chemical analogs of the four DNA nucleotides. These analogs are generally known as the ddNTPs or deoxyribonucleotides.
So, these analogs are missing in the 3’ hydroxyl group but, are generally required for 5’ color for every base, template DNA, and unlabeled dNTPs in a PCR.
As a result, the ddNTPs are randomly incorporated which eventually terminate the chain. This helps in the production of strands of each possible length.
- Next, electrophoresis is used for the separation of extension products. They are again resolved to single nucleotide differences in terms of size.
- After that, the fluorescent labels are mainly used for the detection of the chain terminated fragments.
In this case, each of the colors helps in the identification of the terminating ddNTPs. So, proper analysis helps in the derivation of the sequence of the template DNA strand.
So, the above-discussed ones are considered to be some of the most important things that you should know about the sequencing Sanger.
Also, this particular method of Sanger sequencing has undergone several changes over the last several years. But, despite that, this method is regarded as the most commonly used DNA sequencing technology globally.
Not only that, but this technology of Sanger sequencing is 99.99% accurate. This is also regarded as the gold standard for most of the applications.
Therefore, this method is very useful in both clinical and research. However, the Sanger sequencing is best suited for low and medium-throughput targeted sequencing online projects. Also, in the last decade, this method is used for higher-throughput DNA sequencing technologies.
This is based on fundamentally diverse methods which are known to be the NGS or Next-Generation sequencing. Along with that, this particular technology contributes to offer fast turnaround time i.e. it takes only 4 hours to complete a run.
As a result, both of these parallel technologies have revolutionized the study of molecular biology and genomics.