title: RNA-Seq of Passiflora cincinnata: MIM156
design: Total RNA and Dynabeads Oligo (dT) 25 (Thermo Fisher Scientific, Waltham, MA) were used to isolate mRNA. The resulting mRNA fragments of ~400 nucleotides were converted to double-stranded complementary DNA (cDNA) using random hexamer primers and corresponding enzymes following standard DNA library protocols for sequencing. Briefly, cDNA was end-repaired, phosphorylated, and adenylated. Common TruSeq adapters containing 8-bp indexes (i5 and i7) suitable for Illumina sequencing were then ligated to the adenylated molecules, and the resulting libraries were amplified by 13 cycles of PCR to enrich for properly ligated molecules. The final libraries were quantified using PicoGreen (Thermo Fisher Scientific) and equally combined into a single sample, then sequenced on an Illumina HiSeq X (Illumina Inc., San Diego, CA) instrument. Paired-end reads with an average length of 150 bp were obtained. Library preparation and sequencing were carried out by Rapid Genomics, LLC (Gainesville, FL).
library_name: UFV_114602_P01_WA05