Download A Nanoliter-Scale Nucleic Acid Processor with

Review of:
A Nanoliter-Scale Nucleic Acid
Processor with Parallel Architecture
Tiffany Guo
October 10, 2008
20.309
J. W. Hong, et al. "A nanoliter-scale nucleic
acid processor with parallel architecture,"
Nature Biotech. 22(4): pp. 435-439 (2004)
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
How It Works:
The Components
How it works:
Mixing
How it works:
DNA Isolation by Affinity Column
How it works:
Elution
Parallel Architecture
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
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
expression
patterns
“If a these
specific
combination
of
reflect
the noninvasive
cells
gene expression
patterns
canthat
be
make
up the
bulk
of the tumor
correlated
with
metastic
ability,
mass
and then
that the
however,
suchexpression
analyses
signature
of invasive
tumor cells –
can be predictive
of metastatic
arguably
the motile population
outcome…”
essential for metasasis – might be
masked.”
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)
QUESTIONS?