Name: Juhany M.

Musa

Section: BSMT 1-B

Polymerase Chain Reaction

Description of the Assay

Polymerase Chain Reaction (PCR) is a laboratory technique used to

amplify small amounts of DNA or RNA sequences, producing millions or
billions of copies of a specific segment of DNA in a short period of time.

The PCR process is carried out in a thermal cycler, which repeatedly heats and
cools the reaction mixture containing the template DNA, a pair of primers
(short pieces of single-stranded DNA that complement the ends of the target
sequence), nucleotides (the building blocks of DNA), and a special enzyme
called Taq polymerase.

The three steps of each PCR cycle are:

Denaturation: The reaction mixture is heated to a high temperature (usually

95°C) to separate the double-stranded DNA template into two single strands.

Annealing: The temperature is lowered to allow the primers to bind (anneal) to

their complementary sequences at each end of the target DNA segment.

Extension: The temperature is raised again to the optimal temperature for Taq
polymerase (usually around 72°C), and it extends the primers, adding
nucleotides to synthesize new DNA strands complementary to the template.

After each cycle, the number of DNA molecules doubles, resulting in an

exponential increase in the amount of the target DNA sequence. The number
of cycles can vary, but typically 20 to 40 cycles are performed, resulting in
over a million copies of the target sequence.

PCR has numerous applications in research, diagnosis, and forensic science,

including gene cloning, disease diagnosis, paternity testing, and forensic
identification.
How it works:

Polymerase Chain Reaction (PCR) is a laboratory technique used to

amplify a specific segment of DNA or RNA. The PCR process involves a
series of temperature changes that are carried out in a machine called a
thermal cycler. The steps involved in the PCR process are:

Denaturation: The first step involves heating the reaction mixture to a high
temperature (usually around 95°C) to separate the double-stranded DNA
template into two single strands.

Annealing: The temperature is then lowered to allow short pieces of single-

stranded DNA called primers to bind to complementary sequences at each end
of the target DNA segment.

Extension: The temperature is raised again to the optimal temperature for a

special enzyme called Taq polymerase (usually around 72°C). Taq polymerase
extends the primers by adding nucleotides to synthesize new DNA strands
complementary to the template.

These three steps constitute a single PCR cycle. The number of cycles can
vary, but typically 20 to 40 cycles are performed, resulting in over a million
copies of the target sequence.

The amplified DNA can then be analyzed in various ways, depending on the
purpose of the PCR. For example, the DNA may be sequenced to determine its
nucleotide sequence, or it may be used as a template for further cloning or
manipulation.

PCR is a powerful tool in molecular biology and has numerous applications in

research, diagnosis, and forensic science, including gene cloning, disease
diagnosis, paternity testing, and forensic identification.

Major Breakthrough:

The development of Polymerase Chain Reaction (PCR) by Kary Mullis

in the 1980s is considered a major breakthrough in molecular biology and has
revolutionized the field of biotechnology. Prior to PCR, the amplification of
DNA sequences was a time-consuming and labor-intensive process that
involved cloning the DNA fragments in bacteria or using other complex
methods. PCR allowed researchers to amplify specific segments of DNA
quickly and easily, making it a fundamental tool in many areas of biological
research.

PCR has numerous applications, including DNA sequencing, gene expression

analysis, genetic disease diagnosis, and forensic analysis. It has also played a
crucial role in the development of many biotechnologies, including DNA
fingerprinting, genetic engineering, and gene therapy.

PCR has also had a significant impact on medicine, particularly in the

diagnosis and treatment of infectious diseases. PCR can detect the presence of
viral or bacterial DNA or RNA in patient samples, enabling rapid and accurate
diagnosis of infectious diseases. PCR has also been used to develop new drugs
and vaccines, as well as to monitor the effectiveness of treatments.

Overall, the development of PCR has had a major impact on the field of
molecular biology and has revolutionized many areas of science and medicine.

Closing remarks:

In summary, Polymerase Chain Reaction (PCR) is a laboratory

technique that has had a significant impact on the field of molecular biology
and biotechnology. It allows for the amplification of specific DNA or RNA
sequences, enabling rapid and accurate analysis of genetic information. The
development of PCR has led to many important breakthroughs in medicine,
including the diagnosis and treatment of infectious diseases. PCR is also
widely used in research and has contributed to the development of many
biotechnologies. Overall, PCR is a powerful tool that has revolutionized the
way that scientists study DNA and RNA and has paved the way for many
important discoveries in the field of molecular biology.