|Application:||Gene expression microarray analysis|
|Number of samples:||18|
|Release date:||Jan 29 2008|
|Last update date:||Mar 17 2012|
|Dataset link||profiling of an organ-specific sunflower transcriptoma under abiotic stress (salinity and cold conditions)|
To control biological variation between individuals, three biological samples from the same tissue were pooled on one sample prior to probe preparation. The reference (control) sample consisted of pooled RNA extracted from sunflower seedlings growing under unaltered environmental greenhouse conditions, whereas chilling and salinity samples were RNA extracted from sunflower seedlings growing in greenhouse under those stressed conditions. The RNA (800ng) samples were labeled by using SuperScript Indirect RNA Amplification System Kit (Invitrogen, cat# L1016-02) based on the method designed by Eberwine y col. 1992. Following RNA amplification (with the incorporation of UTP aminoallil), labeled product was achieved by incubating with Cy3 or Cy5 esters in alkaline media. Slides were used in order to quantify the relative expression of ESTs in control and treated leaves by Cy3 and Cy5 hybridization technique Dye-swaps were used to correct for differences in incorporation and fluorescent properties of both dyes, generating a number of 9 slides per experiment (three slides for control and three slides for each treatment) with a total number of 18 slides considering technical replicates.
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BMC Plant Biol