Characterization
Description(s):miRNAs are known to involve in multiple biological functions and abnormal expression is associated to human diseases such as cancers. They represent a promising new class of cancer biomarkers. A number of multiplex methods such as microarray or microbeads assays are commonly used for profiling the expression of multiple miRNAs simultaneously. However these methods need the conversion of the miRNA into a cDNA probe during analysis. Signosis has developed a simple miRNA direct hybridization plate array method, in which total RNA does not need any pre-treatment before hybridization. It is as simple as direct hybridization and detection. More importantly, total 48 miRNAs can be quantitatively analyzed and compared simultaneously with the sample assay.
Applicable Grid:
List of Applicable miRNAs
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | let-7b-5p | miR-17-5p | miR-29a-3p | miR-132-3p | miR-182-5p | miR-206 | let-7b-5p | miR-17-5p | miR-29a-3p | miR-132-3p | miR-182-5p | miR-206 |
| B | miR-1 | miR-18a-5p | miR-34a-5p | miR-133 | miR-192-5p | miR-210-3p | miR-1 | miR-18a-5p | miR-34a-5p | miR-133 | miR-192-5p | miR-210-3p |
| C | miR-7-5p | miR-19a-3p | miR-92-3p | miR-143-3p | miR-194-5p | miR-214-3p | miR-7-5p | miR-19a-3p | miR-92-3p | miR-143-3p | miR-194-5p | miR-214-3p |
| D | miR-9-5p | miR-19b-3p | miR-101a | miR-145-5p | miR-196a-5p | miR-222-3p | miR-9-5p | miR-19b-3p | miR-101a | miR-145-5p | miR-196a-5p | miR-222-3p |
| E | miR-10b-5p | miR-20a-5p | miR-106a-5p | miR-146a-5p | miR-199a-5p | miR-223-3p | miR-10b-5p | miR-20a-5p | miR-106a-5p | miR-146a-5p | miR-199a-5p | miR-223-3p |
| F | miR-15a-5p | miR-21-5p | miR-107 | miR-148a-3p | miR-200 | miR-342-3p | miR-15a-5p | miR-21-5p | miR-107 | miR-148a-3p | miR-200 | miR-342-3p |
| G | miR-15b-5p | miR-24-3p | miR-122a-5p | miR-155-5p | miR-204-5p | miR-372-3p | miR-15b-5p | miR-24-3p | miR-122a-5p | miR-155-5p | miR-204-5p | miR-372-3p |
| H | miR-16-1 | miR-26a-5p | miR-125a-5p | miR-181a-5p | miR-205-5p | U6 | miR-16-1 | miR-26a-5p | miR-125a-5p | miR-181a-5p | miR-205-5p | U6 |
Principle
Signosis’ miRNA direct hybridization plate array is a simple two-step assay; plate hybridization and streptavidin-HRP detection. The plate is pre-coated with an oligo mix, including a pair of unique oligos that hybridize side-by-side to a specific target miRNA and a universal capture oligo and a biotin-labeled oligo. In the assay, total RNA is directly utilized for hybridization. When the target miRNA exists in RNA, it acts as a bridge to bring the biotin-labeled oligo to the capture oligo, which can be detected through streptavidin-HRP conjugate and a chemiluminescent substrate. If there exists no specific miRNA, the biotin-labeled probe will be washed away, leading to no detection. In the plate array, 48 wells are coated with different oligo mixes for different miRNAs. A single 96-well plate allows quantitative measurement and comparison of 48 miRNAs between two samples. U6 RNA is used for normalization.
Data
miRNA direct hybridization plate array I used for analyzing the expression of 48 miRNAs in human skeletal muscle and kidney total RNAs. As indicated in the figure, miR-1 and miRNA 133 are specifically expressed in skeletal muscle, which agrees with published studies.
Literature
View user manualCitations
The role of miR-210, E2F3 and ephrin A3 in angiosarcoma cell proliferation. Satoko Nakashima,Masatoshi Jinnin,Hisashi Kanemaru,Ikko Kajihara,Toshikatsu Igata,Sayo Okamoto,Yukino Tazaki,Shinichi Masuguchi,Satoshi Fukushima,Mamiko Masuzawa,Yasuyuki Amoh,Mikio Masuzawa,Hironobu Ihn, European Journal of Dermatology. 2017 Jul 25, DOI: 10.1684/ejd.2017.3084
Mitochondrial Oxidative Stress-Induced Epigenetic Modifications in Pancreatic Epithelial Cells. Mishra PK, Raghuram GV, Jain D, Jain SK, Khare NK, Pathak N. Int J Toxicol. 2014 Feb 24.
Ankyrin repeat and Single KH domain 1 (ANKHD1) drives renal cancer cell proliferation via binding to and altering a subset of miRNAs. Fragiadaki, Maria, and Martin P. Zeidler. Journal of Biological Chemistry, 2018, doi:10.1074/jbc.ra117.000975.
