GSM2024779: ddcc mutant post-mature green seeds biological replicate 1; Arabidopsis thaliana; RNA-Seq
Collected seeds were quickly frozen in liquid nitrogen and ground to a fine powder using a mortar and pestle. Total RNA was isolated from the powder using the Concert Plant RNA Reagent (Invitrogen, Carlsbad, CA) according to manufacturer’s instructions. Total RNA was treated with RNase-free DNase I (QIAGEN, Valencia, CA) and quantified with Quant-iT™ RiboGreen® RNA reagent (Thermo Fisher Scientific, Grand Island, NY) on a Nanodrop ND-3300. Approximately 25 nanogram of total RNA was amplified with Nugen Ovation RNA-Seq system v2 (Nugen, San Carlos, CA) and quantified with Picogreen on the Nanodrop ND-3300 (Nanodrop Technologies).  One microgram of double-stranded cDNA was used to prepare the Illumina library using the Illumina TruSeq DNA Sample prep kit (Illumina, San Diego, CA). Libraries were quantified using Quant-iT PicoGreen dsDNA Reagent (Thermo Fisher Scientific , Grand Island, NY) and a Nanodrop ND-3300 instrument. Phi-X174 DNA was spiked in to the library by the sequencing facility before cluster formation and sequencing.