Conclusions In this review, an EST database was designed to allow broad characterization with the carnation transcriptome. We detected 17,362 likely basic sequence repeats in 14,291 unigenes and identified transcripts corresponding to genes connected with carotenoid bio synthesis, chlorophyll biosynthesis and degradation, anthocyanin biosynthesis, and ethylene bio synthesis and signaling. This collection of transcripts from carnation might be helpful to the annotation with the forthcoming carnation genome sequence and professional vide a impressive resource for genomics studies in Caryophyllaceae. Procedures Plant supplies and RNA extraction Carnation cultivar Francesco was grown underneath purely natural daylight problems in a green household as described previously, Every single tissue was har vested from 3 plants.
The following plant tissues selleckchem have been made use of. flower bud, flower, younger and adult leaves, and stem, Flowers contained sepals, petals, stamens and pistils. Tissues have been instantly frozen in liquid nitrogen and stored at 80 C. Total RNA was extracted applying the RNeasy Plant Mini Kit, RNA concentration was estimated making use of an ND one thousand spectro photometer and RNA integrity was evaluated employing an Agilent 2100 Bioanalyzer, cDNA library building for 454 sequencing For 454 sequencing, we made a normalized cDNA li brary and also a three cDNA library in cooperation selleck inhibitor with Takara Bio, RNA isolated from every single tissue was mixed in equal proportions in the single pool in an attempt to maximize the diversity of transcriptional units sampled. The Clontech Clever method was employed for cDNA synthesis in the complete RNA.