An oligonucleotide microarray for microRNA expression analysis based on labeling RNA with quantum dot and nanogold probe.
MicroRNAs (miRNAs) play important regulatory roles in animals and plants by targeting mRNAs for cleavage or translational repression. They have diverse expression patterns and might regulate various developmental and physiological processes. Profiling miRNA expression is very helpful for studying biological functions of miRNAs. We report a novel miRNA profiling microarray, in which miRNAs were directly labeled at the 3' terminus with biotin and hybridized with complementary oligo-DNA probes immobilized on glass slides, and subsequently detected by measuring fluorescence of quantum dots labeled with streptavidin bound to miRNAs through streptavidin-biotin interaction. The detection limit of this microarray for miRNA was approximately 0.4 fmol, and the detection dynamic range spanned about 2 orders of magnitude. We made a model microarray to profile 11 miRNAs from leaf and root of rice (Oryza sativa L. ssp. indica) seedlings. The analysis results of the miRNAs had a good reproducibility and were consistent with the northern blot result. To avoid using high-cost detection equipment, colorimetric detection, a method based on nanogold probe coupled with silver enhancement, was also successfully introduced into miRNA profiling microarray detection.
Liang
Ru-Qiang
RQ
Key Laboratory of Proteomics 320 Yue-Yang Road, Shanghai 200031, China.
Li
Wei
W
Li
Yang
Y
Tan
Cui-yan
CY
Li
Jian-Xun
JX
Jin
You-Xin
YX
Ruan
Kang-Cheng
KC
D019921:Biotinylation; D003124:Colorimetry; D020869:Gene Expression Profiling / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D035683:MicroRNAs / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D015335:Molecular Probes / Q000737:chemistry*; D049329:Nanostructures; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012275:Oryza / Q000235:genetics; D045663:Quantum Dots; D018749:RNA, Plant / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012834:Silver / Q000737:chemistry
D035683:MicroRNAs; D015335:Molecular Probes; D018749:RNA, Plant; D012834:Silver; D006046:Gold