Wheat endosperm development

ABSTRACT: Our objective is to describe variations in expression profiles of transcriptional factors during endosperm development, especially during filling phase in which storage proteins are synthesized. Plants from cv. Recital were cultivated under controled conditions. Two biological repetitions were available for grain harvest. at 11 developmental stages (132, 172, 289, 323, 381, 437, 477, 527, 577, 635 and 686 degrees C day). RNA was extracted from all samples of seeds whose embryo was removed.cDNA synthesis, labelling, and hybridization were made according to manufacturer’s protocol (NimbleGen Arrays Users Guide). The SuperScript Double-Stranded cDNA synthesis kit (invitrogen) was used to transform RNA in double-stranded cDNA. The first strand was transcribed with an oligo dT primer and the SuperscriptTM II Reverse Transcriptase. The synthesis of second strand was performed according to the method of Gubler and Hoffman by using RNAse H, DNA ligase, DNA polymerase I. The cDNA was labeled with Cy3 by Random priming by using the NimbleGen One-Color DNA Labeling Kit. Each biological replicate including 11 samples was hybridized on independent microarray. A common sample was placed on both microarrays (stage 323 degrees C day, Biological Repetition 2). The Cy3-labeled cDNA samples were hybridized on microarray by using the NimbleGen Hybridization Kit at 42 degrees c for 18h. Microarrays were washed by using the components of the NimbleGen Wash Buffer Kit and NimbleGen Array Processing Accessories. Then, microarrays were scanned with an Innoscan 900 AL scanner (Innopsys) by using Mapix software. NimbleScan software was used to create signal intensities files from scanned images files. A grid was overlaid on the microarray image to identify the placement of the probes. The software extracts raw data and generates X,Y and signal reports providing the coordinates of each probe and its intensity

ORGANISM(S): Triticum aestivum

SUBMITTER: Catherine Ravel 

PROVIDER: E-MTAB-2952 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress