DNA quantification was performed using the TRIzol DNA extraction kit [1] followed by quantification using a nanodrop. The Trizol reagent simultaneously homogenizes the tissue samples collected at 0, 3, 4, and 5 days of decellularization.. The Trizol lyses the cell membrane and extracts the DNA and RNA. The reagent inhibits DNase activity preserving the integrity during the DNA extraction. Then, chloroform is added to separate the layers of DNA, RNA, and cellular materials. DNA is precipitated using ethanol, and the resulting pellet is resuspended using EDTA. The solubilized pellet is collected for DNA quantification using the NanoDrop.
DNA quantification results, given in ng DNA per mg tissue. Standard for decellularized tissue is < 50 ng/mg tissue. Our results show that 3 days of decellularization is sufficient in removing cellular content.
Page lead: Jonathan Yu
References
[1] "TRIzol Reagent User Guide," Thermo Fisher Scientific, [Online]. Available: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/trizol_reagent.pdf. Accessed: Nov. 19, 2023.