RNAseq of clv1-101 clv2-101 rpk2-5 root segment: BCN 4 dpi (11-day-old seedling), rep3
Developing syncytium (about 8 mm segments) from primary roots of clv1-101 clv2-101 rpk2-5 mutant plants were collected from at 4 dpi (days-post-inoculation)(11-day old seedling) under stereoscope, fixed in ice-cold EAA (ethanol: acetic acid = 3:1) overnight, and embedded in paraffin wax. Samples were sectioned longitudinally on Leica HM 325 rotary microtome, and syncytial cell content were collected on a ArcturusXT Laser Capture Microdissection System (ABI). RNA was isolated with a picoPure RNA Isolation Kit (ThermoFisher Scientific, cat# KIT0204). RNA concentration was determined by Qubit flourometer (Invitrogen) using the Qubit HS RNA assay kit (Invitrogen, Cat# Q32852), and the RNA integrity was checked using the Fragment Analyzer automated electrophoresis system. rRNA was then depleted by Ribominus plant kit (ThermoFisher, cat# A1083808), and ethanol precipitated with glycogen. Library was constructed with Illuminas TruSeq stranded Total RNA Library Prep kit and sequenced on Illumina NextSeq 500.