GSM1655113: SH; Eutrema salsugineum; RNA-Seq
Four biological replicates consisting of five plants each were collected. whole rosettes of 4-week-old plants were frozen in liquid nitrogen and stored at −80°C. Tissue was finely ground in liquid nitrogen using a mortar and pestle. Approximately 800 mg ground tissue was combined with 1 ml of TRIzol reagent (Gibco/BRL Life Technologies, Invitrogen, Carlsbad, CA, USA) and RNA was extracted according to the manufacturer’s instructions. Quality of RNA was estimated by the ratio of absorbance at 260 to 280 nm and 260 to 230 nm by spectrophotometer (DU 730; Beckman Coulter, Inc., Indianapolis, IN, USA) with both ratios between 1.8 and 2.2. RNA was reverse-transcribed into cDNA using the Poly(A) Purist protocol (Ambion, Austin, TX, USA). The cDNA samples were fragmented into 300–500 bp molecules and sequencing libraries were constructed for the 454 GS-FLX instrument (454 Life Sciences, Roche Company, Branford, CT, USA) at the Purdue University Genomics Facility (West Lafayette, IN, USA). One sequencing run was performed. Non-strand-specific protocol was used for 454 library preparation.
GEO Accession
GSM1655113