title: Micro-CRIBI microRNA Platform (mirVana Probe Set V1, Ambion)
status: Public on Jul 14 2014
contact.city: Padova
contact.first: Gerolamo
contact.last: Lanfranchi
contact.email: stefano.cagnin@unipd.it
contact.address: Via U. Bassi, 58/B
contact.department: CRIBI - Biotechnology Center and Biology Department
contact.country: Italy
contact.institute: University of Padova
contact.phone: +39-0498276219
organism: Mus musculus
manufacture_protocol: The mirVana oligo set (Ambion, Austin, TX; Catalog number 1564V1) consists of a collection of about 400 amine-modified DNA oligonucleotides. The probes are 42-46 nucleotides (nt) long, of which an 18-24 nt segment targets a specific known human, mouse, or rat miRNA, and the remaining sequence serves as spacer. Negative control probes were designed using sequences from Escherichia coli; positive control probes (spikes) are sequences from Mytilus galloprovincialis. Each oligonucleotide is spotted by our robotic station Biorobotics Microgrid II in four replicates on a Nexterion Slide E glass slide (Cat No 1066643) provided by SCHOTT. Nexterion Slide E is designed to covalently immobilize amine-modified DNA for microarrays. The deposition is qualitatively assessed by a scan of the spotted slide at 10 micron resolution, using a GSI Lumonics scanner and a protocol that allows the detection of spotted features through the salts derived from the dried solution in which oligonucleotides are dissolved. Microarray slides that meet the quality standards, are washed as recommended by the supplier and the glass slides are stored, ready for use, in the dark under deep vacuum.