1. Review of: A Nanoliter-Scale Nucleic Acid Processor with Parallel Architecture Tiffany Guo October 10, 2008 20.309

2. J. W. Hong, et al. "A nanoliter-scale nucleic acid processor with parallel architecture," Nature Biotech. 22(4): pp. 435-439 (2004)

3. Single Cell Nucleic Acid Isolation Study presence (through DNA) or expression (through RNA) of specific genes – Identify cells of specific genotype for experimentation – Offer clues to differential phenotypes under specific conditions Study transcriptional variation between cells of same genotype

4. How It Works: The Components

5. How it works: Mixing

6. How it works: DNA Isolation by Affinity Column

7. How it works: Elution

8. Parallel Architecture

9. Microfluidics Process requires minimal numbers of cells – Eliminate overnight culturing steps – Study expression on the single cell level – Study expression of slow growing cells more easily

10. Application: Variable Expression of Motility Genes in Tumor Cells W. Wang, et al. “Tumor cells caught in the act of invading: their strategy for enhanced cell motility,” TRENDS in Cell Bio. 15(3): pp. 138-145 (2005) “There is, however, the possibility that these expression patterns reflect the noninvasive cells that make up the bulk of the tumor mass and that the expression signature of invasive tumor cells – arguably the motile population essential for metasasis – might be masked.” “If a specific combination of gene expression patterns can be correlated with metastic ability, however, then such analyses can be predictive of metastatic outcome…”

11. Future Directions: Micro PCR E.A. Ottesen, et al. “Microfluidic Digital PCR Enables Multigene Analysis of Individual Environmental Bacteria, Science, 314: pp. 1464-1467 (2006)

12. QUESTIONS?