miRNA-Seq of Linum usitatissimum: intrusively growing phloem fibers
Intrusively growing phloem fibers (iFIB) were isolated by laser microdissection from longitudinal sections of frozen stem (3rd cm from the apex) obtained by cryosectioning. Total RNA from the iFIB sample (about 500 microdissections obtained from 30 plants) was isolated using RNAqueous-Micro Kit RNA Isolation from Micro-Scale and LCM Samples (Ambion) and library of small RNA were prepared with a NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs, UK) Quality control of the library was performed using an Agilent 2100 Bioanalyzer (Agilent Technologies, USA). Further size selection was performed using Agencourt AMPure XP (Beckman Coulter, USA) to select for 143149 bp fragments. Raw reads from run were filtered to remove adapter sequences, contamination, low-quality reads and any reads shorter than 15nt and longer than 30nt.
SUB5185332
Lus_small_iFIBb_2