Product Name Catalog # Price   Qty
Human miRNA Array IV AP-0009 $549
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Characterization

Description:

Signosis' Human miRNA array IV allows you to profile 100 miRNAs.


Applicable Grid:

List of Applicable miRNAs

miR-138 miR-139 miR-105 miR-130a miR-135a miR-147 miR-152 miR-154-3p miR-184 miR-187
miR-189 miR-211 miR-212 miR-217 miR-22 miR-220 miR-299-3p miR-299-5p miR-301 miR-302a-3p
miR-302a-5p miR-302b-3p miR-302b-5p miR-302c-3p miR-302c-5p miR-302d miR-30d miR-30e-3p miR-30e-5p miR-32
miR-320 miR-323 miR-324-3p miR-324-5p miR-325 miR-326 miR-328 miR-329 miR-33 miR-330
miR-331 miR-335 miR-337 miR-338 miR-339 miR-33b miR-340 miR-345 miR-346 miR-34b
miR-34c miR-361 miR-362 miR-363-3p miR-363-5p miR-365 miR-367 miR-369-3p miR-369-5p miR-370
miR-371 miR-373-5p miR-374 miR-376a-3p miR-376a-5p miR-376b miR-377 miR-378 miR-379 miR-380-3p
miR-380-5p miR-381 miR-382 miR-383 miR-384 miR-409-5p miR-410 miR-411 miR-412 miR-422a
miR-422b miR-423 miR-424 miR-425 miR-425-5p miR-429 miR-432-5p miR-432-3p miR-433 miR-448
miR-449 miR-449b miR-450 miR-451 miR-452-5p miR-452-3p miR-453 miR-455 miR-483 U6

Principle

miRNA is different from large messenger RNA in three aspects; (1) miRNAs are small size molecules with quite a big difference in abundance, (2) mature miRNAs co-exist with their precursor pre-miRNA and pri-miRNA, which only differ in length, and (3) many miRNAs are very closely related in sequences, such as isoforms, only differing by one or a few nucleotides. Therefore, the conventional mircoarray technologies cannot directly be applied to analyzing these molecules. A number of miRNA microarray products are commercially available, but they are either tedious in requiring pre-isolation of microRNA, lack the discriminative power to differentiate between isoforms, or are not sensitive enough to monitor low abundant miRNAs.

In our array assay, each miRNA molecule is targeted by two oligos that hybridize with the target miRNA to form a RNA/DNA duplex. When the sequences are perfectly matched, the oligos are aligned with the miRNA and the joint can be ligated by DNA ligase (figure 1). A single nucleotide difference among miRNAs will block either the hybridization or the ligation; Thus, miRNA isoforms can be differentiated. Due to the small size of miRNA, the hybrid might not be stable; Therefore, we introduce the stacking sequences. By extending these two oligos along with their complementary oligos, the stability is increased. Once the pair of oligos is ligated, the ligated molecules are subjected to linear amplification via T7 transcription into RNA in the presence of biotin-UTP, which are used as probes for array hybridization. To differentiate each isoform, we assigned unique tag sequences to the ligation oligos, so that single nucleotide differences are converted into unique tag sequences. In this way, each isoform can be easily distinguished by array hybridization.

The assay procedure includes three steps: (1) mix the miRNA with provided oligos to form miRNA/oligo hybrids; (2) select the hybrids and remove free oligos, and ligate miRNA-directed pairing of oligos to become a single DNA; and (3) amplify the ligated DNA with T7 transcription.


Data



Human miRNA array IV analysis of miRNA expression in human liver and brain tissue.

Literature

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