GSM5745889: Roots, 24hpt, NaCl, B; Arabidopsis thaliana; RNA-Seq
Root systems were collected at 24 hours and 7 days post treatment and flash frozen in liquid nitrogen. Two biological replicates were collected for each time point with each replicate consisting of at least 50 root systems. Root tissue was homogenized with Invitrogen PureLink Plant RNA Reagent (Thermo Fisher Scientific, Waltham, MA, USA) to isolate nucleic acids, and DNA contamination was removed by treating the samples with Ambion TURBO-DNA-free ( Thermo Fisher Scientific, Waltham, MA, USA) per the manufacturer's protocol. RNA quantity was determined with a Nanodrop spectrophotometer and RNA quality was determined with an RNA nanochip on the Agilent 2100 bioanalyzer system Reverse transcription, fragmentation, end repair, adapter ligation, and PCR enrichment were conducted with the NEBNext Ultra II DNA Library Prep Kit (New England Biolabs, Whitby, ON, Canada) per the manufacturer's protocols. Library size distribution was determined with a high sensitivity DNA chip on the Agilent 2100 bioanalyzer system. Library concentration and molarity was determined with the NEBNext Library Quant Kit (New England Biolabs, Whitby, ON, Canada) in conjunction with PerfeCTa qPCR ToughMix (QuantaBio, Beverly, MA, USA) and PicoGreen fluorospectrophotometry. Libraries were sent to Genome Québec (Montréal, QC, Canada) for 150 bp single-end sequencing on the Illumnia HiSeq 4000 platform.