title: RNA-seq of Black Hybrid at Stage 2 - Rep 3
design: Meristems were collected by cutting and shaving plant stolks with a blade to uncover the floral meristems which were measured in mm and frozen in liquid nitrogen immeiatly after isolation. Total RNA was isolated from meristem tissues using the Quick-DNA/RNA Plant/Seed Miniprep Kit (Zymo Research Corporation, Irvine, California, USA) according to kit instructions. Sample purity and quantification was read on a DropQuant spectrophotometer and run on an Agilent Fragment Analyzer using a high sensitivity RNA kit. Only samples with a RIN score greater than 7 were used for sequencing. Sample libraries were sequenced using the Illumina NovaSeq X Plus platform by AgriLife Genomics and Bioinformatics Services (Texas A&M University). Sequence cluster identification, quality prefiltering, base calling and uncertainty assessment were done in real time using the Illumina Sequencing System Software Suite v1.2.0 with default parameter settings. Sequencer .cbcl basecall files were demultiplexed and formatted into .fastq files using the Illumina bcl2fastq v2.20.0.422 demultiplexing software.