GSM9427926: Full-thickness skin wound, non-diabetic, rep 3; Sus scrofa; RNA-Seq
Excised tissue from biopsy punches was stored in RNAlater™ Stabilization Solution (Invitrogen) at -80 °C until the time of RNA isolation. Total RNA was extracted by lysing tissue in TRIzol™ Reagent (Invitrogen) using a TissueLyser II (QIAGEN). The tissue homogenate was phase-separated with chloroform, and the aqueous RNA phase was further purified using a RNeasy Mini Kit (QIAGEN) per the manufacturer's recommendations. Library construction was completed by Novogene Co, Ltd. - Messenger RNAwaspurified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. Then the second strand cDNA was synthesized using dUTP, instead of dTTP. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection.