Project name: Solanum lycopersicum strain:Heinz 1706

Description: Transcriptome analysis of 7 tissues of commercial tomato (S. lycopersicum cv MoneyMaker) and its wild red-fruited ancestor (S. pimpinellifolium LA0722) genotypes performed to assess expression level of tomato transcriptome and to aid whole genome annotation. Sequencing of fruit at 3 different developmental stages will help to assess gene regulation through ripening. Overall design: Plants of S. lycopersicum (cv MoneyMaker) or S. pimpinellifolium (LA0722) were grown in soil-filled pots in greenhouse under controlled conditions in Long day (16 hours light, 8 hours dark). RNA was extracted from different tissues according to published methods(López-Gómez and Gomez-Lim, 1992). cDNA was produced according to the SMART cDNA PCR synthesis kit (Clontech) but using the SuperScriptII Reverse Transcriptase (Invitrogen), their quantity and quality were evaluated respectively by Nanodrop spectrophotometer and Agilent Bioanalyzer (DNA12000 assay). Tissues used for cDNA synthesis in S. lycopersicum were: root, stem, leaf, flower and fruit at 3 different maturation stages: mature green, breaker and ripe. S. pimpinellofolium tissues used for synthesis were leaf and berry ripe. cDNA libraries were produced following the 454 Titanium Rapid Library Preparation protocol instructions, then fixed to micro beads and clonally amplified by emulsion PCR (emPCR Method Manual-Lib-L). The beads have been enriched to keep only those carrying identical PCR products, and then each sample has been loaded on a half picotiter plate to obtain at least 350000 reads with a medium length of 400bp. Each sample was sequenced on a half picotiterplate.

Data type: Transcriptome or Gene expression

Sample scope: Multiisolate

Last updated: 2013-07-26