title: MPIIB Ambion mirVana
status: Public on Jan 01 2010
contact.city: Berlin
contact.first: Hans-Joachim
contact.last: Mollenkopf
contact.email: mollenkopf@mpiib-berlin.mpg.de
contact.address: Charitéplatz 1
contact.country: Germany
contact.institute: Max-Planck-Institute for Infection Biology
contact.phone: +49 30 28460 482
organism: Homo sapiens
  Mus musculus
  Rattus norvegicus
manufacture_protocol: Probes for 200 miRNA (mirVANA Probe Set, lot 124K02) were obtained at 500 pmol each as a collection of amine-modified DNA oligonucleotides targeting a comprehensive selection of human, mouse, and rat miRNAs (Ambion Europe Ltd., Huntingdon, UK). The probes were 42–46 nucleotides (nt) long consisting of a spacer and 18–24 nt specific miRNA segments. They were suspended at a final concentration of 100 µM in Nexterion spot2 buffer (Schott AG, Mainz, Germany) and printed by a BioRobotics MicroGrid II robot (Genomic Solutions Ltd., Huntingdon, UK) onto glass slides coated with poly-L-lysine  at 50-55% humidity and room temperature using MicroSpot 2500 quill pins (Genomic Solutions Ltd.). Each slide contained 4 identical replicates with 4 subgrids. Oligonucleotides were crosslinked to the support by UV irradiation with a total energy of 240mJ UV and blocked with succinic anhydride and 1-methyl-2-pyrrolidone in borate buffer (Fluka, Deisendorf,