Direct multiplexed measurement of gene expression with color-coded probe pairs.
We describe a technology, the NanoString nCounter gene expression system, which captures and counts individual mRNA transcripts. Advantages over existing platforms include direct measurement of mRNA expression levels without enzymatic reactions or bias, sensitivity coupled with high multiplex capability, and digital readout. Experiments performed on 509 human genes yielded a replicate correlation coefficient of 0.999, a detection limit between 0.1 fM and 0.5 fM, and a linear dynamic range of over 500-fold. Comparison of the NanoString nCounter gene expression system with microarrays and TaqMan PCR demonstrated that the nCounter system is more sensitive than microarrays and similar in sensitivity to real-time PCR. Finally, a comparison of transcript levels for 21 genes across seven samples measured by the nCounter system and SYBR Green real-time PCR demonstrated similar patterns of gene expression at all transcript levels.
Geiss
Gary K
GK
NanoString Technologies Inc., 201 Elliott Ave West, Suite 300, Seattle, Washington 98119, USA. ggeiss@nanostring.com
Bumgarner
Roger E
RE
Birditt
Brian
B
Dahl
Timothy
T
Dowidar
Naeem
N
Dunaway
Dwayne L
DL
Fell
H Perry
HP
Ferree
Sean
S
George
Renee D
RD
Grogan
Tammy
T
James
Jeffrey J
JJ
Maysuria
Malini
M
Mitton
Jeffrey D
JD
Oliveri
Paola
P
Osborn
Jennifer L
JL
Peng
Tao
T
Ratcliffe
Amber L
AL
Webster
Philippa J
PJ
Davidson
Eric H
EH
Hood
Leroy
L
Dimitrov
Krassen
K
D002460:Cell Line; D003116:Color; D015342:DNA Probes / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D017930:Genes, Reporter; D006801:Humans; D007091:Image Processing, Computer-Assisted; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D017955:Poliovirus; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D015342:DNA Probes; D012333:RNA, Messenger