--- title: "Finishing extractions" author: "Kathleen Durkin" date: "2025-07-11" categories: ["SIFP_2025"] format: html bibliography: ../../../references.bib --- Completed *E. tourneforti* extractions ### Qiagen DNEasy Extraction Protocol (after digestion) 2. Add **200uL Buffer AL**. Mix thoroughly by vortexing, and spin down the sides. 3. Add **200ul 95% ethanol**. Mix thoroughly by vortexing, and spin down the sides. 4. Pipet mixture onto a labelled DNEasy Mini Spin Column placed in a 2mL collection tube. Centrifuge \@ \>= 8,000 RPM for 1 min. Discard the flow through and collection tube. 5. Place column in a new 2mL collection tube. Add **500uL Buffer AW1**. Centrifuge at \>= 8,000 RPM for 1 min. Discard the flow through and collection tube. 6. Place column in a new 2mL collection tube. Add **500uL Buffer AW2**. Centrifuge at 14,000 RPM for 3 min. Discard the flow through and collection tube. 7. Transfer the spin column to a new, labelled 1.5mL or 2mL tube. 8. Elute the DNA by adding **50uL Buffer AE** to the center of the spin column membrane. Incubate at room temperature for 1 min. Centrifuge 1 min. at \>= 8,000 RPM. 9. Repeat step 8, for a [final elution volume of 100uL]{.underline}. Completed Round 1 extractions at 3:00pm, July 11, 2025. Stored in fridge. Completed Round 2 extractions at 10:00am, July 12, 2025. Stored in fridge.