RNA-seq of skin wound implantation with 3D-bioprinted constructs
RNA-seq of skin wound implantation with 3D-bioprinted constructs
Skin wound tissues were collected from adult female C57BL/6J mice at day 7 post-surgery. The wound area was excised, rinsed in cold PBS, and snap-frozen in liquid nitrogen. Samples were stored at −80 °C until RNA extraction. Total RNA was extracted from frozen wound tissues using TRIzol reagent (Invitrogen, USA) according to the manufacturer's instructions. RNA quantity and integrity were evaluated by Qubit 3.0 fluorometer (Thermo Fisher Scientific) and Agilent 4200 TapeStation (Agilent Technologies). Samples with RIN ≥ 7 were used for library preparation. RNA-seq libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB, USA). Poly(A)+ mRNA was purified, fragmented, and reverse-transcribed to cDNA. After end-repair, adapter ligation, and PCR amplification, the libraries were quantified by Qubit and assessed with the Agilent 4200 system.