"RNA expression profiling at the single molecule level"
, in Genome Research, Seite(n) 1041-1045, 2006, ISSN: 1549-5469
RNA expression profiling at the single molecule level
Sprache des Titels:
We developed a microarray platform for PCR amplification-independent expression profiling of minute samples. A
novel scanning system combined with specialized biochips enables detection down to individual fluorescent
oligonucleotide molecules specifically hybridized to their complementary sequence over the entire biochip surface of
cm2 size. A detection limit of 1.3 fM target oligonucleotide concentration—corresponding to only 39,000 molecules
in the sample solution—and a dynamic range of 4.7 orders of magnitude have been achieved. The applicability of
the system to PCR amplification-independent gene-expression profiling of minute samples was demonstrated by
complex hybridization of cDNA derived from the equivalent of only 104 cells, which matches results obtained in
ensemble studies on large samples. By counting each hybridized molecule on the microarray, the method is
insusceptible to gene-specific variations of the labeling, thereby representing a principle advance to conventional
ensemble-based microarray analysis.