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Introduction to Analysis of Genomic Data Using RLecture10: Introduction to Microarray Technology
(Chapter 5 in Gondro’s book )
Dr. Yen-Yi Ho ([email protected])
March 21, 2018
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Objectives of Lecture 10
I Types of ArraysI Expression Microarrays
I General Experiment ProtocolI Hybridization, Target, Probe, Probe SetI Commercial MicroarraysI Common Issues
I aCGH
I ChIP-chip
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Microarrays
I Microarrays (or just arrays) allow the simultaneousmeasurement of the mRNA of thousands of genes (geneexpression)
I Why “micro”? Measuring things on a small scale means lessmRNA is necessary to be extracted
I The simultaneous measurement is important, so that variousfactors/conditions are all common to the array, and relativegene expression can be measured with less variability
I Still microarray? Microarray is still widely used because oflower costs, easier experimental procedure and moreestablished analysis methods.
I Similar problems are presented in newer technologies such asRNA-seq, and similar statistical approaches can be applied.
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Microarrays
Microarrays can be used for many purpose including:
I genotyping:
SNP arrays
I measure gene expression
I determine DNA copy number: aCGH
I determining transcription factor binding sties: Chip-chip
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Microarrays
Microarrays can be used for many purpose including:
I genotyping: SNP arrays
I measure gene expression
I determine DNA copy number:
aCGH
I determining transcription factor binding sties: Chip-chip
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Microarrays
Microarrays can be used for many purpose including:
I genotyping: SNP arrays
I measure gene expression
I determine DNA copy number: aCGH
I determining transcription factor binding sties:
Chip-chip
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Microarrays
Microarrays can be used for many purpose including:
I genotyping: SNP arrays
I measure gene expression
I determine DNA copy number: aCGH
I determining transcription factor binding sties: Chip-chip
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Co$t
MicorarrayAffymetrix Cost: $425/per arrayIllumina BeadChip Cost: $ 80 ∼ $250 /per strip × 12 = $960 ∼$3,000 /chip
Next-Generation SequencingIllumina ∼$2,000 per samples (for 1-7 samples)
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On Type of Microarray Experimental Procedure
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Target
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Probes
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On Type of Microarray Experimental Procedure
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Platforms that dominate market
I Affymetrix
I Agilent
I Illumina
I NimbleGen
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Affymetrix GeneChip R©
source: Affymatrix
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Affymetrix GeneChip R©
source: Affymatrix17/35
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Affymetrix GeneChip R©
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Probe Set
A probe set is a collection of probes designed to interrogate agiven sequence.
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PM versus MM
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Affymetrix GeneChip R© Experiment Protocol
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Illumina BeadArrays
• Each silica bead is 3 µ in diameter
• 700,000 copies of same probe sequence attached to each bead
• May have more than one bead for a particular gene
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Illumina BeadArrays
• RefSeq BeadChip (left) 8 arrays per chip, 1 strip= 1 array
• Whole Genome (right) 6 arrays per chip, 2 strip= 1 array
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Commons Issues
I Background: print-tip, plate, print-order, spatial effects
I Between arrays: batches, plates, cross platform comparison,experiment protocols
I Within arrays: background noise, intensity dependent effect
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Spatial Effect
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Spatial Effect
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Microarrays
Microarrays can be used for many purpose including:
I genotyping: SNP arrays
I measure gene expression
I determine DNA copy number: aCGH
I determining transcription factor binding sties: Chip-chip
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Array-comparative genomic hybridization (aCGH)
Nature Education1(1). Microarray-based Comparative Genomic Hybridization (aCGH)
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array CGH
Brian and Development 31(6):629:633.
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Microarrays
Microarrays can be used for many purpose including:
I genotyping: SNP arrays
I measure gene expression
I determine DNA copy number: aCGH
I determining transcription factor binding sties: Chip-Chip
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ChIP-chipChromatin immunoprecipitation (”ChIP”) with microarraytechnology (”chip”)
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ChIP-chip
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Readings
Primer to Analysis of Genomic Data Using R (page 163 – 200).
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