title: DOME_Dsv.halophilus_900_v1
status: Public on Dec 22 2010
contact.city: Vienna
contact.first: Doris
contact.last: Steger
contact.state: Vienna
contact.address: Althanstrasse 14
contact.department: Department of Microbial Ecology
contact.country: Austria
contact.institute: University of Vienna
organism: Pseudodesulfovibrio halophilus
manufacture_protocol: A published 16S rRNA-targeted oligonucleotide probe, DVHO831 (S-*-Dv.h.o-0831-a-A-18, 5’-GAA CCC AAC GGC CCG ACA-3’), synthesized by MWG Biotech (Ebersberg, Germany), was used for microarray spotting. The 5’ end of the oligonucleotide was tailed with 12 dTTPs (T-spacer) and the terminal dTTP was aminated to enable covalent coupling of the oligonucleotide to aldehyde group-coated VSS-25 glass slides (CEL Associates). The probe concentration was adjusted to 50 pmol/µl in 50% dimethyl sulfoxide. Microarrays were printed with a single split pin (MicroSpot 2500, Zinsser Analytic GmbH) using a BioRobotics MicroGrid spotter (Genomics Solutions) at 20°C and greater than 50% humidity. Square arrays of identical spots (approximate diameter 150 µm) were produced (30 x 30 spots). Spotted microarrays were incubated overnight at room temperature in a wet chamber and processed with sodium borohydride, as described previously.