Vitis vinifera cv. Corvina (clone 48) samples were collected from a 7-year-old vineyard (45° 27’ 17” N, 11° 03’ 14” E, Montorio, Verona province, Italy) during 2008/2009 growing seasons. The vineyard is at 130 m. above sea level (a.s.l.), planted on a soil with the following composition: 36% sand, 36% say and 28% silt. The replacement cane Guyot rows are north-south oriented and 41B in used as rootstock.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted from ~100 mg of liquid-nitrogen-grinded tissue using the Spectrum™ Plant Total RNA kit (Sigma-Aldrich, St. Louis, MO) following the manufacturer’s protocol. For berry flesh, senescing leaf and woody stem, it was necessary to isolate RNA from ~400 mg of grinded material, while for berry pericarp and skin, seed, rachis, root, latent and winter bud, from ~200 mg. Lithium Chloride precipitation treatment was applied to some of the total RNA samples (from winter bud, from seed and rachis at vèraison and mid-ripening, from woody stem) to remove 230wavelenght-absorbant contaminants. Briefly, LiCl solution was mixed with total RNA solution to a final concentration of 2.5 M; after over-night incubation at 4°C, a cold centrifugation and ethanol-70% washing step, total RNA was resuspended in water.
Label
Cy3
Label protocol
cDNA synthesis and labeling reactions were performed according to the NimbleGen Arrays User’s Guide (V 3.2).
Hybridization protocol
Hybridization and washing procedures were performed according to the NimbleGen Arrays User’s Guide (V 3.2).
Scan protocol
Each microarray was scanned using a Axon GenePix 4400 A at 532 nm (Cy-3 absorption peak) and GenePix Pro7 software (Molecular Devices, Sunnyvale, CA, USA) according to the manufacturers' instructions.
Description
flesh_phw_III_1_431659A09.pair
Data processing
Images were analyzed using NimbleScan v2.5 software (Roche), background correction and standard RMA normalization were selected.
