title: GSM8789919: PBa3D4; Populus balsamifera; RNA-Seq
library_construction_protocol: Total RNA was extracted from frozen ground leaf bud powder using the modified cetyl trimethyl ammonium bromide method for polyphenol-rich plant tissue described by Muoki et al. (2012). MagJET enrichment kit (ThermoScientific) was used to purify mRNA from crude RNA. The mRNA was fragmented by heating the samples at 94℃ for 5 minutes. To make cDNA libraries with adaptors NEBNext ultra RNA Library Prep Kit for Illumina (NEB) was used. GeneJET NGS Cleanup Kit (ThermoScientific) was used to clean the extra adaptors.  Primers for RNA-Seq were added to cDNA using NEBNext Multiplex Oligo kit (NEB). MagJET NGS Cleanup and Size Selection kit (ThermoScientific) was used to select DNA fragments around 350bp. Twelve libraries with same concentration (250 ng/ul) each were pooled and sent to University of British Columbia Sequencing and Bioinformatics Consortium (Vancouver, BC, Canada).