Maxwell 16 mdx as3000

Total DNA was isolated to perform a polymerase chain reaction (PCR) and determine mitochondrial DNA damage. DNA was isolated from renal biopsies of eight controls subjects and twelve sepsis-AKI patients. First, sections of 5 μm thickness were cut from the renal biopsies. Samples were pretreated with 500 µL collagenase V (300 U/mL; Sigma-Aldrich, Darmstadt, Germany), incubated at 37 °C for 3 h and vortexed every 30 min. Subsequently, 500 µL RPMI (ThermoFisher, Paisly, UK) was added, followed by centrifugation for 10 min at 20,000 G at room temperature. Next, Rapid Sample Concentrator (RSC) blood DNA kit (Promega, Madison, USA) was used to isolate DNA from the pellet using Maxwell 16 MDx AS3000 (Promega), according to the manufacturer’s protocol. DNA from HUVECs was isolated with Nucleospin DNA kit (MACHEREY–NAGEL, GmbH & Co. KG, Germany), according to manufacturer’s protocol.