XXX_100_105_C10_R48-415_pt1
For each sample, the first expanding leaf from a side branch was harvested in the field and immediately frozen in individual tubes placed in an ethanol-dry ice bath. Frozen tissues were pulverized using the Qiagen TissueLyser, and total RNA extracted using the Norgen Plant Total RNA Kit (catalog #25800) along with the Norgen in-column DNAse I Kit (catalog #25710) and following the manufacturers instructions. RNA quality was assessed using a NanoDrop and 1.5% agarose gels. RNA concentrations were quantified using Invitrogen Qubit RNA BR Assay Kit (catalog #Q10210). RNA-Seq libraries were created using the KAPA mRNA Hyper Prep (catalog #KK8581) following the manufacturers instructions and ligated with in-house 8-bp dual index adapters. Libraries were enriched for 7-9 cycles and quantified using a plate fluorometer with SYBR Green I. Libraries were pooled in equimolar amounts based on SYBR Green I values and sequenced on Illumina HiSeq4000/NovaSeq6000 sequencers. The sequencing was carried out at the DNA Technologies and Expression Analysis Cores at the UC Davis Genome Center, supported by NIH Shared Instrumentation Grant 1S10OD010786-01.