What is DNA Sequencing?

DNA sequencing is a technique used to,

well… sequence DNA: it allows us to find the
base pair sequence on a strand of DNA.
Although there are many types of DNA
sequencing techniques, the most common
type is Sanger sequencing which will be
discussed in more depth. However, newer
next-generation-sequencing techniques are
being developed and used to make this
process much faster and economical.

Where is DNA Sequencing Used?

DNA sequencing has huge applications. One such
Figure 2: Diagram of Sanger Sequencing process
being genome sequencing. In fact, in the Human
Genome Project, a modified version of sanger
sequencing was the main technique used. Further,
genetic analysis with DNA profiling – a branch of
DNA sequencing – can give lead to better medical
outcomes for genetic diseases. Similarly, the list
goes on and on: from forensics to evolution, there
are a vast number of uses for this technique.

DNA
The Process of DNA
Sequencing – Sanger
Sequencing?
Sequencing
The process of DNA sequencing
starts similarly to PCR
(polymerase chain reaction).
Initially, the DNA mixture is
subjected to high temperature
(~95˚C) to denature the DNA
strands. After denaturation, the
mixture is cooled (~60˚C) to
facilitate the binding of primer
strands to the DNA strands.
Subsequently, a slight increase in
temperature (~70˚C) brings the
enzyme to its optimal Figure 2: normal nucleotide (dNTP) vs probe-acting nucleotide (ddNTP)
temperature, allowing it to extend
the primer. However, during this
extension step, the polymerase
Whenever a ddNTP is added, it chemically stops the extension from continuing on
utilizes a combination of regular
nucleotides and dideoxy- that strand. As these steps are repeated, by chance, a strand will have a ddNTP at
nucleotides (ddNTPs). each specific nucleotide location. All the DNA of different lengths can be isolated
and run through a capillary gel electrophoresis setup. Due to shorter fragments
(which stopped at the first nucleotide) falling through first, we can determine the
order of bases via which ddNTP has been added. Typically, a laser is used to detect
the specific dye, and the DNA sequence can be determined.