title: Food Safety Potato cDNA Microarray 1.0
status: Public on Feb 01 2009
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
  Solanum phureja
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 12 μl 5 x SSC and used for spotting. Microarrays were spotted on sylilated slides (CELAssociates, USA) at 19°C and 55% relative humidity using Microgrid instrument (Apogent technologies, USA) in 6 blocks containing 26 x 26 spots. 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 seven times in each block of the array as well as a Salmonella gene fragment as a negative control. 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 free aldehyde groups were blocked with NaBH4.