title: Food Safety Potato cDNA Microarray 4K 2.0
status: Public on Jan 31 2012
contact.city: Wageningen
contact.first: Jeroen
contact.last: van Dijk
contact.email: jeroen.vandijk@wur.nl
contact.address: Bornsesteeg 45
contact.country: Netherlands
contact.institute: RIKILT, institute of food safety
contact.phone: +31317480256
organism: Solanum tuberosum
manufacture_protocol: PCR fragments were purified through Sephadex G-50 columns (Sigma-Aldrich, USA) and precipitated by overnight incubation with of 1/10 V 3M sodium acetate (pH 5.2) and 1 V of isopropanol. The pellets were washed with 70% ethanol, air-dried, redissolved in 20 μl 50% DMSO-MiliQ and used for spotting. Microarrays were spotted on UltraGAPS slides (Corning) in 64 blocks containing 16 x 16 spots. Two complete microarrays were spotted per slide. As positive controls three different luciferase fragments corresponding to the 5', middle and 3' part of the cDNA, as well as the entire luciferase cDNA were spotted across the array as well as a Salmonella gene fragment as a negative control.A total of 264 controls were included. Microarrays were dried and stored at room temperature until use. Spotting quality was assessed for two of the spotted arrays in the series by staining the slides with 0.2 μM cyanine nucleic acid dye POPO-3 (Molecular probes, USA). Prior to hybridisation DNA was UV-crosslinked to the glass surface with 1500 mJ.