Tools Used in Genetic
Engineering
Biotechnology 8
Genetic Engineering
The process of using recombinant
DNA (rDNA) technology to alter
the genetic makeup of an
organism.
Traditionally,
humans
have
manipulated genomes indirectly
by controlling and selecting
offspring with desired traits.
The Different Tools Used
in Genetic Engineering
Polymerase Chain Reaction (PCR)
Restriction Enzymes (Molecular Scissors)
Gel Electrophoresis
DNA Ligase
Polymerases
Prokaryotic Host
Eukaryotic Host
Selection of Small Self-Replicating DNA
Methods to Select Transgenic Organisms
Polymerase Chain
Reaction (PCR)
Polymerase Chain Reaction (PCR)
The technique is used in molecular biology
to amplify a single copy or a few copies of a
segment of DNA across several orders of
magnitude,
generating
thousands
to
millions of copies of a particular DNA
sequence.
Developed in 1983 Kary Mullis,
PCR is now common technique used
in clinical research laboratories for
various applications. (Javed, 2017)
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Polymerase Chain Reaction (PCR)
PCR has three main stages, which
involve a process of heating and
cooling:
https://www.yourgenome.org
Step 1, denaturing: the double-stranded template
DNA is heated, which separates it into two single
strands.
https://www.yourgenome.org
Step 2, annealing: the temperature is lowered
to enable the DNA primers to attach to the
template DNA.
https://www.yourgenome.org
Step 3, extending: the temperature is raised again and the
new strand of DNA is made by the Taq polymerase enzyme.
https://www.yourgenome.org
These three stages are repeated 20-40 times,
doubling the number of DNA copies each time. This
is called thermal cycling, carried out by a machine.
It’s carried out by a machine called a thermal
cycler. Depending on the speed of the machine,
PCR can be performed in under and hour or up to a
Thermal Cycler
few hours.
https://www.escolifesciences.com
After PCR has been completed, a method called
electrophoresis can be used to check the quantity
and size of the DNA fragments produced.
https://www.genome.gov
Electrophoresis
https://www.yourgenome.org