Illumina RNA-Seq of White Spruce PG653 Tween Treated Cortical resin duct cells
Isolation of RNA for laser microdissected cells was done using the Ambion RNAqueous¨ -Micro Kit according to the manufacturer instructions. Briefly, frozen cells in lysis buffer were thawed on ice and transferred from the cap to the bottom of the tube by pipetting. Lysis buffer was added to a final volume of 100 ľl and the sample was incubated at 42ĄC for 30 min. Each tube received 3ul of LCM additive and was mixed by briefly vortexing. Tubes containing cells from the same tree and cell-type were combined together and received 0.5 volumes of 100% ethanol to recover only large RNA species. The lysate was passed through the kit RNA binding column, washed and eluted according to the manufacturer instructions. RNA integrity and quality were assessed in a Nano chip in a 2100 Bioanalyzer (Agilent Technologies, USA). RNA was concentrated using the GeneJET RNA Cleanup and Concentration Micro Kit (Thermo Scientific, USA).