Bio 302 Remaining Practicals

DNA fingerprinting:
A laboratory genetic technique or method to identify individuals using bodily samples such
as blood, saliva, or hair is referred to as DNA fingerprinting. Or we can define DNA
fingerprinting as, A DNA test to establish a link or relation between two person or living
organisms by analyzing their STR and VNTRs is known as DNA fingerprinting.
The technique of DNA fingerprinting changed the era of identification, characterization, and
classification of organisms. Notably, approximately 99% of our DNA (deoxyribose nucleic
acid) is similar. A 0.1% difference is sufficient to make someone so unique. We are using this
0.1% portion for DNA profiling which is often known as DNA fingerprinting.
Satellites DNA:
Mainly two types of satellite regions are present in the human genome based on their repeat
sequence nature: minisatellite and microsatellites. Minisatellite region contains repeated
DNA sequences of 10 to 60 bp. 5 to 50 repeats of it are present in our genome. For example,
VNTRs. microsatellites are smaller than minisatellites. It’s 1-6 bp long and repeated 5 to 10
times in a genome. For example, STR and SSR.
Restriction Fragment Length Polymorphism (RFLP):
Restriction fragment length polymorphism (RFLP) is a technique invented in 1984 by the
English scientist Alec Jeffreys during research into hereditary diseases. It is used for the
analysis of unique patterns in DNA fragments in order to genetically differentiate between
organisms – these patterns are called Variable Number of Tandem Repeats (VNTRs). Genetic
polymorphism is defined as the inherited genetic differences among individuals in over 1% of
normal population. The RFLP technique exploits these differences in DNA sequences to
recognize and study both intraspecies and interspecies variation.
Principle:
Restriction endonucleases are enzymes that cut lengthy DNA into short pieces. Each
restriction endonuclease targets different nucleotide sequences in a DNA strand and therefore
cuts at different sites. The distance between the cleavage sites of a certain restriction
endonuclease differs between individuals. Hence, the length of the DNA fragments produced
by a restriction endonuclease will differ across both individual organisms and species.
Competent Cells
Competent cells are bacteria that have been treated to increase their ability to uptake foreign
DNA molecules. This heightened DNA uptake efficiency enables efficient transformation, a
process by which foreign DNA is introduced into the cells. Competent cells play a crucial
role in molecular biology research, as they facilitate the introduction of recombinant DNA,
enabling genetic manipulation, gene expression studies, and the production of proteins of
interest in biotechnology applications.
Analysis of proteins by SDS-PAGE:
SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) is a widely used
laboratory technique for separating and analyzing proteins based on their size. In this method,
proteins are denatured by the detergent SDS, which imparts a negative charge proportional to
their mass. The proteins are then loaded onto a polyacrylamide gel and subjected to an
electric field. Smaller proteins migrate faster through the gel, leading to their separation into
distinct bands. This technique is essential for protein characterization, quantification, and
purification in various biological and biochemical research applications.
SDS (Sodium Dodecyl Sulfate): SDS is a detergent commonly used in molecular biology and
biochemistry. It denatures proteins by binding to them and unfolding their three-dimensional
structures. SDS also imparts a uniform negative charge to proteins, enabling them to migrate
primarily based on their size during electrophoresis.
PAGE (Polyacrylamide Gel Electrophoresis): PAGE is a technique used to separate
molecules, such as proteins or nucleic acids, based on their size and charge. A
polyacrylamide gel matrix is used as a medium, and an electric field is applied, causing
charged molecules to migrate through the gel. The smaller molecules move faster, leading to
their separation into distinct bands, allowing for analysis and purification.

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