Wastewater DNA/RNA
Extraction Kit
Quick Start Guide
Optional
To avoid taking pelleted or floating material:
For processing 10ml, 15ml should be spun down to collect clear supernatants, leaving behind 5ml in the tube.
For higher biomass samples, or to obtain a full biomass profile with wastewater samples, a bead beating step is required during the lysis step. Add 1 ml of activated sludge in bead beating tube, then proceed directly to step 3.
- Add 100l of Concentration Beads (mix well before use) to 10ml wastewater sample, then invert 5 times to mix thoroughly. Incubate for 10 mins. At the 5 min mark, invert 3 times to mix the beads.
Note: Store the beads at 4°C.
- Place sample on 15ml magnetic rack to capture beads, then discard supernatant.
- Add 400yl of Lysis Buffer. Resuspend the beads by pipetting up-down. Transfer mixture to a clean 2ml centrifuge tube. Mix for 5 mins using vortexer at maximum speed.
- Use a 2ml magnetic rack to capture the beads. Avoiding the pellet, transfer up to 400l of the supematant into a clean 2ml microcentrifuge tube.
- Add 600l of Binding Buffer. Add 30ul of Binding Beads. Vortex for 10-20s, and incubate for 5 mins.
- Place tube on magnetic rack to capture, wait 1 min then discard supernatant.
- Add 600l of Wash Buffer #1 to the tube and vortex for 10-20s, Wait 1 min then place the tube on the magnetic rack to capture. Wait 1 min then discard supernatant.
- Add 600ul of Wash Buffer #2 to the tube and vortex for 10-20s. Place the tube on a magnetic rack to capture. Wait 1 min then discard supernatant.
- Add 100yl of Elution buffer to tube and mix briefly. Incubate at 65°C for 10 mins, Place the tube on a magnetic rack to capture.
- Wait 1 min then transfer supernatant to clean 2ml microcentrifuge tube.
Note: For increased yield perform elution twice with 50yl of buffer.
Wastewater DNA/RNA Extraction Kit III” W010 0-12