title: RNA-seq of rat tissues with chronic osteomyelitis
design: RNA-seq of rat tissues with chronic osteomyelitis
library_construction_protocol: In this study, five femoral tissue samples were collected from the chronic osteomyelitis model group and the control group each, resulting in a total of ten samples. Total RNA extraction using Total RNA extraction Kit (DP431, TIANGEN, China). The quality and quantity of the purified RNA were determined by measuring the absorbance at 260nm/280nm(A260/A280) using Ultrafine spectrophotometer (N50 touch, IMPLEN, Germany). The integrity of RNA was further verified by 1.0% agarose gel electrophoresis For each sample, 500 ng of total RNA was used for cDNA synthesis via a strand-switching reverse transcription approach. The resulting cDNA was then amplified by PCR, followed by barcoding with the Native Barcode kit (SQK-NBD114.96) and purification using Ampure XP Beads. Finally, 300 fmol of the resulting library was loaded onto a PromethION flow cell (vR10.4.4) for sequencing.