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MICROARRAY
R. PERUMAL
2016603110
M. Sc., FLORICULTURE & LANDSCAPING
WHAT IS ARRAY???
 An array is an orderly arrangement of samples
where matching of known and unknown DNA
samples is done based on base pairing rules.
 An array experiment makes use of common assay
systems such as microplates or standard blotting
membranes.
HISTORICAL BACKGROUND
 Microarray technology evolved from
Southern blotting
 The concept of DNA microarrays began in
the mid 1980s.
 Mark Schena was proclaimed as the
“Father of Microarray Technology”
Mark Schena
MICROARRAY
 Microarrays are sets of miniaturized chemical
reaction areas that may be used to test DNA
fragments, antibodies, or proteins
 Each reaction area or spot is having immobilised
target which is hybridised with complimentary probe
present in the testing sample
PRINCIPLE
 The core principle behind microarrays is hybridization
between two DNA strands.
 Fluorescent labeled target sequences that bind to a
probe sequence generate a signal that depends on the
strength of the hybridization determined by the number
of paired bases.
“Microarray” has become a general term, there are many
types now
 DNA microarrays
 Protein microarrays
 Transfection microarrays
 Antibody microarray
 Tissue microarray
 Chemical compound microarray
DNA MICROARRAY
 A DNA microarray is a collection of microscopic DNA spots on solid
surface. Each spot contains pico moles of a specific DNA sequence,
known as probes or reporters
 Used for detection of polymorphisms and mutations in genomic DNA
 Commonly known as DNA chip, DNA arrays or biochips
 Bio chip is a small rectangular solid surface that is made of glass or
silicone
TYPES OF DNA MICROARRAY
i. Glass cDNA microarrays which involves the micro spotting of pre-
fabricated cDNA fragments on a glass slide.
ii. High-density oligonucleotide microarrays often referred to as a
"chip" which involves in situ oligonucleotide synthesis.
STEPS
I. Collect Samples
II. Isolate mRNA
III. Create Labelled DNA
IV. Hybridization
V. Microarray Scanner
VI. Analyze Data
EQUIPMENT'S
HYBRIDIZATION
Hybridization for 16-20 hrs at 45-60⁰ C
IMAGE ACQUISITION
 Unbound material is washed away
and the sample hybridized to each
element is visualized by
fluorescence detection.
 Fluorescence emission from the
microarray is converted into a
digital output for each dye, and is
stored as separate image files.
DNA MICROARRAY SCANNER
Fluorescent intensity is measured
THE COLOURS OF A MICROARRAY
 GREEN represents Control DNA, where either DNA or cDNA
derived from normal tissue is hybridized to the target DNA.
 RED represents Sample DNA, where either DNA or cDNA is
derived from diseased tissue hybridized to the target DNA.
 YELLOW represents a combination of Control and Sample
DNA, where both hybridized equally to the target DNA.
 BLACK represents areas where neither the Control nor
Sample DNA hybridized to the target DNA.
APPLICATIONS OF MICROARRAY TECHNOLOGY
CONCLUSION
Microarrays measure the concentration of mRNA
Several assumptions are made in the progress of analysis
Biological
Numerical
Experimental conditions are important!
Different distances matrics combined with different clustering
algorithms can leed to completely different results
THANK YOU

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Microarray

  • 1. MICROARRAY R. PERUMAL 2016603110 M. Sc., FLORICULTURE & LANDSCAPING
  • 2. WHAT IS ARRAY???  An array is an orderly arrangement of samples where matching of known and unknown DNA samples is done based on base pairing rules.  An array experiment makes use of common assay systems such as microplates or standard blotting membranes.
  • 3. HISTORICAL BACKGROUND  Microarray technology evolved from Southern blotting  The concept of DNA microarrays began in the mid 1980s.  Mark Schena was proclaimed as the “Father of Microarray Technology” Mark Schena
  • 4. MICROARRAY  Microarrays are sets of miniaturized chemical reaction areas that may be used to test DNA fragments, antibodies, or proteins  Each reaction area or spot is having immobilised target which is hybridised with complimentary probe present in the testing sample
  • 5. PRINCIPLE  The core principle behind microarrays is hybridization between two DNA strands.  Fluorescent labeled target sequences that bind to a probe sequence generate a signal that depends on the strength of the hybridization determined by the number of paired bases.
  • 6. “Microarray” has become a general term, there are many types now  DNA microarrays  Protein microarrays  Transfection microarrays  Antibody microarray  Tissue microarray  Chemical compound microarray
  • 7. DNA MICROARRAY  A DNA microarray is a collection of microscopic DNA spots on solid surface. Each spot contains pico moles of a specific DNA sequence, known as probes or reporters  Used for detection of polymorphisms and mutations in genomic DNA  Commonly known as DNA chip, DNA arrays or biochips  Bio chip is a small rectangular solid surface that is made of glass or silicone
  • 8. TYPES OF DNA MICROARRAY i. Glass cDNA microarrays which involves the micro spotting of pre- fabricated cDNA fragments on a glass slide. ii. High-density oligonucleotide microarrays often referred to as a "chip" which involves in situ oligonucleotide synthesis.
  • 9. STEPS I. Collect Samples II. Isolate mRNA III. Create Labelled DNA IV. Hybridization V. Microarray Scanner VI. Analyze Data
  • 12.
  • 13. IMAGE ACQUISITION  Unbound material is washed away and the sample hybridized to each element is visualized by fluorescence detection.  Fluorescence emission from the microarray is converted into a digital output for each dye, and is stored as separate image files.
  • 14. DNA MICROARRAY SCANNER Fluorescent intensity is measured
  • 15. THE COLOURS OF A MICROARRAY  GREEN represents Control DNA, where either DNA or cDNA derived from normal tissue is hybridized to the target DNA.  RED represents Sample DNA, where either DNA or cDNA is derived from diseased tissue hybridized to the target DNA.  YELLOW represents a combination of Control and Sample DNA, where both hybridized equally to the target DNA.  BLACK represents areas where neither the Control nor Sample DNA hybridized to the target DNA.
  • 16.
  • 17.
  • 19. CONCLUSION Microarrays measure the concentration of mRNA Several assumptions are made in the progress of analysis Biological Numerical Experimental conditions are important! Different distances matrics combined with different clustering algorithms can leed to completely different results