Genotyping Microarray

What are Microarrays?

Microarrays are laboratory tools used to detect the expression of thousands of genes at the same time. They consist of a collection of slides printed with DNA fragments, known as probes, which are used to hybridize a cDNA sample. The probes are DNA fragments that are either synthetized in situ or spotted on the array using photolithographic masks.

Microarray Support Construction

Oligonucleotides are synthetized in situ and used to create probes. This process is called photolithographic synthesis. The probes are then fixed to the array, which supports the thousands of microscopic spots containing the DNA probes.

Sample Preparation

In order to prepare the sample for microarray analysis, mRNA is isolated from the tissue. The isolated mRNA is then used to make a labeled DNA copy. In the case of cancer samples, both tumor and normal tissue samples are collected and processed in the same way.

Hybridization

In the hybridization step, the labeled cDNA molecules bind to the DNA probes on the slide. This binding occurs due to the property of complementary nucleic acid sequences.

sequences to pair with eachother

High number of complementary basepairs means tighter non-covalentbonding between strands

After washing off non-specificbonding sequences, only stronglypaired strands will remain hybridized

HYBRIDIZATION

HYBRIDIZATION

HYBRIDIZATION

DATA ANALYSIS

Why do we obtain yellow?

ElectromagneticSpectrum

Emission Wavele Frequength ncy

Radiowavelength Microwaves

Infrared

Ultraviolet

Excited X-rayswavelength p Gamma-raysp<

Why do we obtain yellow?

Wavele Frequength ncy IntensityThreshold

It needs small support and few reagents

Rapid and automatic method

Why It can screen many genesmicroarray It allows direct and parallel comparisons? between different genes

We can obtain great amount ofinformations in a short time

It’s not toxic or radioactive

Applications

ONCOLOGY PHARMACOGENE MICROBIOLOGY

TICSONCOLOGYDISEASEDISCOVERY

• Compare differentpatterns of geneexpression

• Map geneticabnormalities in a widerange of tumors

Microarrays for Cancer Diagnosis and Classification

1. Analysis of cancer genomes through microarrays and next-generation sequencing (Li Liu, Alex Yick-Lun So, Jian-Bing Fan)
2. ONCOLOGY DRUG DISEASE DISCOVERY
• Detect treatments
• Compare different patterns of gene expression according to patient
• Confirm mechanisms of action of drugs
• Map genetic abnormalities in a wide range of tumors
• Predict chemoresistance
3. Microarrays for Cancer Diagnosis and Classification (Ainhoa Perez-Diez, Andrey Morgun, and Natalia Shulzhenko)
4. Analysis of cancer genomes through microarrays and next-generation sequencing (Li Liu, Alex Yick-Lun So, Jian-Bing Fan)
5. ONCOLOGY DRUG DISEASE DIAGNOSES AND DISCOVERY PROGNOSIS
• Progress of disease
• Detect treatments
• Compare different patterns of gene expression according to patient
• Predict therapies or relapse risk
• Compare expression
• Confirm mechanisms
• Map genetic profile of abnormalities in a wide range of tumors

1. Predict chemorange of tumors cancersresistance
2. Microarrays for Cancer Diagnosis and Classification (Ainhoa Perez-Diez, Andrey Morgun, and Natalia Shulzhenko.)
3. Analysis of cancer genomes through microarrays and next-generation sequencing (Li Liu, Alex Yick-Lun So, Jian-Bing Fan)
4. PHARMACOGENETICS
5. PERSONALIZED DECIDE DRUGMEDICINE TREATMENTS AND METABOLISM
6. THERAPIES
7. DNA microarrays and pharmacogenomics (Rolando Meloni, Olfa Khalfallah, Nicole Faucon Biguet)
8. MICROBIOLOGY
9. COMPARATIVE Provide useful information for the identification of
10. GENOMICS AND virulence factors, exploration of molecular phylogeny,
11. GENOTYPING development of vaccines
12. DNA Microarrays – Techniques and Applications in Microbial Systems (T.Majtán, G. Bukovská, J. Timko)
13. MICROBIOLOGY
14. COMPARATIVE Provide useful information for the identification of
15. GENOMICS AND virulence factors, exploration of molecular phylogeny,
16. GENOTYPING development of

DETERMINATION OF Virulence gene expression can be monitored by growing the

VIRULENCE FACTORS pathogens in vivo and comparing them with the expression of

OF MICROBIALPATHOGENS genes under in vitro conditions

DNA Microarrays – Techniques and Applications in Microbial Systems (T.Majtán, G. Bukovská, J. Timko)