Microarray of long oligonucleotide

LONG OLIGONUCLEOTIDE
MICROARRAY

Microarray:
 A microarray (also commonly known as
biochip) is a collection of microscopic spots
of nucleotide attached to a solid surface.
 Each spot contains picomoles (10−12 moles)
of a specific DNA sequence, known as
probes (or oligos).
 Each known gene or “probe” occupies a
particular “spot” on the chip
 Fluorescently labeled target sequences that
bind to a probe sequence generate a signal.

Oligonucleotide:
 Oligonucleotides are short nucleic acid
polymers used in research, genetic testing
and forensics.
 Oligonucleotides are usually made up of
13 to 25 nucleotides [DNA /RNA]and are
designed to hybridize specifically to DNA
or RNA sequences.

 The term oligonucleotide is derived from
the Greek “oligo,” which means few or
small. The length of the oligonucleotide is
usually denoted by the term “mer,”which
is Greek for “part.”
 Oligonucleotides are used as probes for
detecting specific sequences that are
complementary to the oligonucleotides.

Oligonucleotide Microarray:
 In oligonucleotide microarrays , the
probes are short sequences designed
to match parts of the sequence of
interest.
 Oligonucleotide arrays are produced
by printing short oligonucleotide
sequences designed to represent a
single gene by synthesizing this
sequence directly onto the array
surface

Principle of Oligonucleotide
Microarray
 The core principle behind microarrays is
hybridization.
 Samples are labelled using fluorescent
dyes.
 At least two samples are hybridized to
chip.
 Complementary nucleic acid sequences
get pair via hydrogen bonds.
 Washing off of non-specific bonding
sequences and scanner viewing.

Procedure:
 Preparation of the sample[DNA/RNA]
 Preparation of radiolabelled
complementary nucleotide
 hybridization
 Arraying in the chip
 Viewing under scanner
 Predicting results.

Making of Substrate:
 The substrate is the important concept
in the microarray technology
 The substrate for arraying the
hybridized oligonucleotide may be a
GLASS SILDE or metal coated slides
 Nowadays the BIOCHIPS are only
used as the substrate

Biochips:
• Commercially available
• Biochip acts as the
substrate material.
• A microchip are
designed to predict the
function of hybridized
biological sample.
• The number of spots
can be performed is
1600 spots per Cm2

Sample collection and
purification
The isolated
bacteria was then
moved for
isolation of
nucleotide[either
DNA/RNA]
Bacteria was
isolated by pure
culture technique
[or]
Other human
nucleotides can
also be taken

Labelling:
• The extracted oligonucleotide sample
was to be mixed with the Radiolabelled
complementary oligonucleotide
• And polymerisation was done

Hybridization:
 The control sample was mixed with
experimental sample by robots.
radiolabelled

Arraying:
Done by robots on biochips. 10-12 picomoles

Predicting the results:
 Laser scanners
 Excellent spatial resolution
 Good sensitivity, but can bleach fluorochromes
 Still rather slow
 CCD scanners
 Low resolution
 Sensitivity, easily adjustable (exposure time)
 Faster and cheaper than lasers

 Green- control nucleotide
 Red- sample nucleotide
 Yellow-combination of control and
sample.

Result in oligonucleotide
Microarray:

Overall process:
Example: cancer oligonucleotide

Applications:
 Gene expression analysis and study
of genes
 Transcription factor binding analysis
 Drug discovery for infectious diseases
 Oligonucleotide microarrays are
medically important and are much
useful to carry out genomic
researches.

Microarray of long oligonucleotide

Microarray of long oligonucleotide

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Microarray of long oligonucleotide