DNA Microarray Analysis Jay Edneil C. Olivar
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DNA Microarray Analysis
Jay Edneil C. Olivar
DNA Microarray
DNA Microarray
The Result
DNA Microarray: The Experiment
Healthy Cell Cancer Cellvs.
Aliquot of RNA
DNA Microarray: mRNA isolation
Column with poly-T beads
DNA Microarray: the FUN part
Polymerization of labeled cDNA
Green: HealthyRed: Cancer
DNA Microarray: ResultsNormal
Cancer
Combined
DNA Microarray: Results
Upregulated in cancer cells
Downregulated in cancer cells
Expressed in both cells
Not Expressed in both cells
Limits to Microarray analysis
Measure of gene expression is solely based on the production of an mRNA
For more information
• Visit: http://learn.genetics.utah.edu/content/labs/microarray/
Advantages and Disadvantages
Advantages DisadvantagesSimultaneous analysis of multiple genes
Confusing to first time users
High throughput Result not always reproducible
Allows comparison of gene expression (healthy vs. non-healthy)
Gene expression is dictated by qualitative data
Can categorize diseases into subgroups
Still too expensive
Objective
•To determine ANEUPLOIDY RATES and INCIDENCE OF MOSAICSIM in good-quality human preimplantation embryos using an array-based technique
Aneuploidy and Mosaicism
•Aneuploidy•Condition wherein chromosome
number is abnormal•Mosaicism•A condition wherein an individual has
two or more cell populations with a different chromosomal makeup
Methodology
Computational Analysis
Array Hybridization
Whole genome extraction; Amplification
10 embryos (70 blastomeres) from 6 successful in vitro fertilization was subject to testing
Microarray in the Experiment
22 x 22
22 x 22
22 x 22
22 x 22
Printed twiceRepresents about 7744 genes
1. Plated genes were designed to contain all possible aberrations
2. Detection of aberrations was done using the CIRCULAR BINARY SEGMENTATION (CBS) method
3. DNA probing done through Cy3 and Cy5 labeled probes
Results
Embryos0
10
20
30
40
50
60
70
80
Normal Mosaic
Blastomeres0
10
20
30
40
50
60
NormalWith Chromosomal Abnormalities
Conclusion and Implication
•High rate of aneuploidy and mosaicism in excellent quality embryos from cycles with a high implantation rate WARRANTS FURTHER RESEARCH on the origin and significance of chromosomal abnormalities in human preimplantation embryos.
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