Acuderm acu punch

Using an Acuderm Acu-Punch (Fisher Scientific, Pittsburg, PA) tool, eight 4.0 mm biopsy punches were sampled per placenta. Each sample was stored individually in a sterile, DNase- and RNase-free 2.0 mL Nalgene long-term storage cryogenic tube (Nalgene, Rochester, NY) with 1mL of TRIzol preservative solution (Fisher Scientific, Pittsburg, PA). The samples were snap frozen in a dry ice and isopropanol slurry and then transferred to a −80°C freezer. RNA was extracted either through the VANTAGE core using a modified Fibrous tissue protocol with the QIAsymphony RNA kit (Qiagen, Hilden, Germany) or in a collaborating VUMC laboratory using a modified Qiagen’s miRNeasy Mini Kit Protocol (Qiagen, Hilden, Germany; see Sup Mat 1).

Using an Acuderm Acu-Punch (Fisher Scientific, Pittsburg, PA) tool, eight 8.0 mm biopsy punches were sampled per placenta. Each sample was stored individually in a sterile, DNase- and RNase-free 2.0 mL Nalgene long-term storage cryogenic tube (Nalgene, Rochester, NY) with 1 mL of DNAgard preservative solution (Sigma-Aldrich, St. Louis, MO). The samples were snap frozen in a dry ice and isopropanol slurry. Samples were stored at −80°C before DNA was extracted in the Vanderbilt Technologies for Advanced Genomics (VANTAGE) core (see Sup Mat 1), and quantified using a NanoDrop 8000 Spectrophotometer (Fisher Scientific, Pittsburg, PA).