Review on Application of DNA microarray
Introduction:
Definition of DNA microarray:
It is solid supports, usually of glass or silicon, upon which DNA is attached in an
organized grid fashion. Uses of DNA microarray in versatile technology it used for parallel
gene expression analysis from 1000 of genes of known and unknown function and it is also
used for DNA homology analysis for detecting polymorphism and mutation.
History of DNA microarray:
This type of microarray technology evolved from southern blotting technology. They
are used for gene expression profiling and it was reported in 1995. A complete eukaryotic
genome on a microarray was published in 1997.
Principle of DNA microarray:
DNA microarray consists of thousands of wells or spots, each well or spot contain
DNA or particular gene which acts as probe. Whole spot contains DNA or particular gene
represents the whole genome of an organisms.
Main principle of DNA microarray is Hybridization between two DNA strands and
the presence of one genomic or cDNA sequence in 1 lakh per molecule and this sequence can
be screened in a single hybridization.
Steps involved in DNA microarray
a) Preparation of DNA chip.
b) Extraction of total RNA/DNA:
c) Involves conversion of mRNA into cDNA and label fluorescently.
d) Mix labelled cDNA together for hybridization of cDNA to array.
e) Wash un-hybridized cDNA.
f) Analysis images.
Ex: To known the expression of genes in disease and normal cell, then applied the prepared
cDNA and spot DNA. Then it is measured under fluorescent light.
Results: Red colour shows the more gene expression which is present in normal cell it is
mainly due to up regulation of that gene.
Green colour shows the less gene expression which is present in diseased cell it is
mainly due to down regulation of that gene.
Yellow colour shows gene product presents in both normal and diseased cells.
Technique used in DNA microarray:
a) Photolithography
b) Robot spotter
c) Ink jet method
Types of microarray: Classification based on 3 criteria’s
a) Size of the probes used: It may be complementary DNA array or oligonucleotide
array.
b) Manufacturing method: It consist deposition and in-situ synthesis.
c) Number of samples that can be simultaneously profiled on one array: It involves
single channel array and multiple channel arrays.
Applications of DNA microarray:
a) Gene discovery and expression profiling
b) Discovery of drugs
c) Disease diagnostics
d) Toxicological research
e) Food quality
f) Microbial detection
Advantage of DNA microarray:
a) It helps in the detection of expression of thousands of genes at a timws
b) It provides data for thousands of genes
c) One experiment instead of many
d) Fast and easy to obtain results
e) Huge step closes to discovering and cure diseases and cancer.
Limitation of DNA microarray:
a) Array provide an indirect measure of relative concentration.
b) For complex mammalian genomes, it is often difficult to design array.
c) DNA array can only detect sequences that the array was designed to detect.
Future line of prospect:
Use unbiased method to directly measure all the DNA/RNA species presents in a
particular sample.
The advent of next generation sequencing technologies combined the rapid decreases
in the cost of sequencing.
It provides a strategy for researchers studying gene expression, methylation, and
mutation studies.
Conclusion:
Identifying of thousands genes at a time, DNA microarray helps in identification of
new drug designing and play role in toxicological research and it provide the strategy to
understand epigenetic gene expression studies.