title: miRXplore TM Microarray (836)
status: Public on Nov 28 2012
contact.city: Bergisch Gladbach
contact.first: Silvia
contact.last: Rueberg
contact.email: silvia@miltenyibiotec.de
contact.address: Friedrich-Ebert-Str. 68
contact.department: Genomic Services
contact.country: Germany
contact.institute: Miltenyi Biotec GmbH
organism: Homo sapiens
  Mus musculus
  Rattus norvegicus
description: The miRXplore TM Microarray is composed of 5668 DNA oligonucleotides with a sequence being reverse complement to the respective mature miRNA or control RNA, representing a total of 1320 miRNA sequences and 95 controls with all probes spotted in quadruplicates. The 1320 miRNA sequences covered sequence-verified miRNAs of the species human (735), mouse (587) and rat (429), as well as viral miRNAs (150) as deposited in the miRBase sequence database version 10.0. The 97 positive and negative control probes were chosen as follows. Three sets of controls, miRControl 1 (four sequences used to assess efficiency and sensitivity of labelling), miRControl 2 (single sequence used to assess efficiency of small RNA enrichment) and miRControl 3 (43 sequences used to normalize signal intensities across different samples and arrays) were artificial sequences chosen not to be present in any of the human, mouse, rat or viral genomes. A fourth set of positive controls presented 36 positive small but non-miRNA sequences including probes e.g., for 5.1 sRNA, tRNAs, and U6 RNAs. A fifth set of oligonucleotides included 13 negative controls like different single or double mismatch, antisense, or scrambled miRNA sequences. Microarrays were manufactured by Miltenyi Biotec. Briefly, 1.2 nl of 10 µM HPLC purified, 5’ amino-modified DNA oligonucleotides (Metabion) were spotted on aminopropyltrimethoxysilane- / phenylendiisothiocyanate-coated glass slides in quadruplicates using a customized piezoelectric spotter.
manufacture_protocol: Defined 200-400 bp fragments of selected cDNAs were generated by RT-PCR, cloned and sequence-verified. Amplified and purified cDNA inserts were spotted in quadruplicate on PIQOR slides using a proprietary piezoelectric spotter. The PIQOR slides were coated in a 2-step process generating a surface which covalently binds DNA molecules.