BIO 351
DNA profiling or fingerprinting
Every living thing is composed of segments of deoxyribonucleotides called genes. When
these genes code for the production of amino acids, they are called exons. Scattered between
exons are introns which do not code for the production of amino acids but are however
important in regulation of gene expression. The totality of all the genes in an individual is
referred to as its genome. One inherits the genome vertically from both parents as pairs of
chromosomes (23 pairs: half from each parent) during fertilisation. Each gene occupies a
locus on these chromosomes.
As humans are descendants of a common ancestor and do not readily engage in horizontal
gene transfer, any two individuals will have genomes that are 99.9% identical with only a
difference of 0.1%. This ‘little’ 0.1% accounts for sequence differences in over 300 million
base pairs. These differences are usually due to mutations. So, any two individuals will differ
in certain regions of the chromosome and as such no two individuals will have exactly the
same genome (even identical twins). An individual’s unique genomic characteristics is that
individual’s DNA fingerprint or profile. And as we know, no two individuals have an
identical fingerprint.
Regions of the chromosome known as variable number of tandem repeats (VNTRs) are
important tools in DNA profiling. These are repeating sequences of DNA arranged in tandem
(one after another). These repeating sequences differ from one individual to another in
number of repetitions and are thus said to be variable. VNTRs vary in length of nucleotides
or base pairs, and thus affect the overall length of chromosomes. When the VNTRs are less
than 10 base pairs (typically 1-6 base pairs), they are called microsatellites; and are called
minisatellites when the sequence length is greater than 10 base pairs. These VNTRs are found
in over 1000 locations on the chromosome and are unique to each individual due to their high
propensity to mutation and hence sequence variation.
In addition to VNTRs, a type of polymorphism (sequence differences) known as restriction
fragment length polymorphism (RFLP) is also essential in DNA profiling. RFLPs are due to
changes in the number of restriction sites found in an individual’s chromosome. These
restriction sites are specific sequences of nucleotides recognised by restriction enzymes.
These recognition enzymes cut DNA at their individual restriction sites. Due to mutation,
most individuals have variations in their number of restriction sites and thus the number of
times their DNA is cut by a particular restriction enzyme and the length of the resulting cut
DNA fragments. VNTRs are important sources of RFLPs since most restriction sites are
found within minisatellites and microsatellites. Together, both VNTRs and RFLPs are
important tools for DNA profiling due to their variation among individuals.
Often (especially in forensic and paternity/maternity investigations) there comes a need to use
each individual’s unique DNA profile as a tool for identification. In forensic studies, experts
use DNA profiling in identifying criminals, while paternity/maternity tests involve
confirmation of a child’s parents. Determination of the number and nature of VNTRs and
RFLPs aid in these investigations.
Techniques in DNA profiling
Two main techniques are used in DNA profiling: an older technique based on RFLP and
southern blotting and a newer PCR-based technique.
Steps in RFLP and Southern blotting technique
Gene tracking techniques for prenatal disease diagnosis.
