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Microarray (Gene Chip) Technology

Microarray (Gene Chip) Technology. PART 1. Part 2. Part 3. Part 4. Features – Parallelism • Thousands of genes simultaneously – Miniaturization • Small chip size – Multiplexing • Multiple samples at the same time – Automation • Chip manufacturing • Reagents. Part 1.

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Microarray (Gene Chip) Technology

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  1. Microarray (Gene Chip) Technology

  2. PART 1 Part 2 Part 3 Part 4

  3. Features – Parallelism • Thousands of genes simultaneously – Miniaturization • Small chip size – Multiplexing • Multiple samples at the same time – Automation • Chip manufacturing • Reagents

  4. Part 1 PART 2 Part 3 Part 4

  5. Step Step Step

  6. Step 2 Step 1 Step 3 Step 2 Step 3

  7. Microarray Fabrication • Fabrication via Printing (cDNA chip) • DNA sequence stuck to glass substrate • DNA solution pre-synthesized in the lab • Fabrication In Situ(Affymetrix chip) • Sequence “built” • Photolithographic techniques use light to release capping chemicals • 365 nm light allows 20-m resolution

  8. Fabrication via printing cDNA chip Fabrication In Situ: Affymetrix chip

  9. Microarray (Gene Chip) Technology

  10. Microarray Features – Parallelism • Thousands of genes simultaneously – Miniaturization • Small chip size – Multiplexing • Multiple samples at the same time – Automation • Chip manufacturing • Reagents

  11. Afftmetrix chip

  12. cDNA chip

  13. cDNA chip

  14. Fabrication via printing cDNA chip Fabrication In Situ: Affymetrix chip

  15. Microarray experiment and data acquisition

  16. Poly-A mRNA 10% Biotin-labeled Uracil Antisense cRNA cDNA IVT AAAA Cells L L L L L L Fragment (heat, Mg2+) Labeled fragments Hybridize Wash/stain Scan Affymetrix microarray process

  17. L L L L L L L L L L L L Hybridization and Staining Biotin Labeled cRNA GeneChip Hybridized Array + + SAPE Streptavidin- phycoerythrin

  18. The Result • A light source scans the array, causing the dyes to fluoresce • The glow is picked up by a sensor and is used to determine the relative abundance of the RNA • This information must be processed to determine the level of activity for each expressed gene

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