Extraction of Total Nucleic Acid from Environmental Samples for the Detection of Bacterial and Viral Targets v1

This procedure describes a modified extraction procedure to isolate both DNA and RNA from wastewater samples using the QIAamp Fast DNA Stool MiniKit. The sample undergoes a lysate preparation process and includes mechanical disruption (bead beating), removal of inhibitors, purification and elution of DNA and RNA using spin columns. Extrinsic controls SPC and MS2 are added to each sample during the lysate preparation to evaluate extraction and amplification efficiency. The extracted total nucleic acid (TNA) is then stored at -80°C for testing. To rule out contamination during the extraction process, a blank is also processed through the complete protocol each day extractions are performed. Briefly, the procedure comprises the following steps: a. Lysis of and separation of impurities from sample pellets in InhibitEX buffer. b. Purification of DNA and RNA on QIAamp Mini spin columns.