Transcriptome sequencing of Asclepias syriaca - fc1048-lane4-index06
"We mechanically homogenized fresh frozen leaves on dry ice in a FastPrep-24 bead mill (MP Bio). We added 1.5 ml of cold extraction buffer to the ground tissue, homogenized the tissue and pelleted cellular debris. The supernatant was incubated overnight at 4?C, RNA was pelleted by centrifugation and cleaned. We prepared an RNA-Seq library using 2 ?g total RNA, followed by ribosomal RNA subtraction (RiboMinus?; Life Technologies), and standard TruSeq reagents (Illumina Inc.) modified to enable strand-specific sequencing by dUTP incorporation (Parkhomchuk et al., 2009)."