Ceramic bead tubes kit

Sample preparation has been described elsewhere [13 (link)]. Shortly, biopsies were allowed to thaw at room temperature (RT) under a cytostatic hood. The biopsies were transferred into labeled 2 mL vials and kept at +4 °C in a fridge before lyophilization. Then, vials were placed into a Speedvac device (S-Concentrator, BA-VC-300H; H. Saur, Laborbe-darf, Reutlingen, Germany) and centrifuged under vacuum overnight (1,000 rpm; 100 mbar) at RT. The dry pellets were weighed on a high accuracy scale (R180D; Sartorius, Germany) for later normalization. Then, the dry pellets were rehydrated with 1.5 mL of sterile distilled water (Ampuwa, Fresenius KABI, Homburg, Germany) and homogenized using a homogenizer (TissueLyser LT; QIAGEN GmbH, Hilden, Germany). Shortly after, the sample material and ceramic beads were placed together into 2 mL ceramic tubes (Ceramic Bead Tubes Kit; QIAGEN GmbH, Hilden, Germany) and shaken in a vertical position (50 Hz, 3,000 oscillations/min) for 1 h at RT. Then, the tubes were placed into an ultrasounication device (Elmasonic S30H; Singen, Germany) for 10 min at RT. Finally, the tubes were mixed on a vortex mixer for 30 s, centrifuged for 10 min (5417R, 9,000 rpm; Eppendorf, Hamburg, Germany) at RT and stored at −80 °C.

Biopsies were allowed to thaw at room temperature (RT) under a cytostatic hood. The biopsies were transferred into labeled 2-ml vials and kept at + 4° C in a fridge before lyophilization. Then, vials were placed into in a Speedvac device (S-Concentrator, BA-VC-300H; H. Saur, Laborbedarf, Reutlingen, Germany) and centrifuged under vacuum overnight (1000 rpm; 100 mbar) at RT. The dry pellets were weighed on a high accuracy scale (R180D; Sartorius, Germany) for later normalization. Then, the dry pellets were rehydrated with 1.5 ml of sterile distilled water (Ampuwa, Fresenius KABI, Homburg, Germany) and homogenized using a homogenizer (TissueLyser LT; QIAGEN GmbH, Hilden, Germany). Shortly after, the sample material and ceramic beads were placed together into 2 ml ceramic tubes (Ceramic Bead Tubes Kit; QIAGEN GmbH, Hilden, Germany) and shaken in a vertical position (50 Hz, 3000 oscillations/min) for 1 h at RT. Then, the tubes were placed into an ultrasounication device (Elmasonic S30H; Singen, Germany) for 10 min at RT. Finally, the tubes were mixed on a vortex mixer for 30 s, centrifuged for 10 min (5417R, 9000 rpm; Eppendorf, Hamburg, Germany) at RT and stored at − 80 °C.