DOH Workshop Part 2: Promega Pronex protocol

Resuspend the Pronex beads by vortexing for 0h 0m 10sor longer.

Note

Into your extracted DNA tube pipette 80µLof Pronex beads and mix into the sample by slowly pipetting 10 times.

Leave at Room temperature for 0h 10m 0s.

Place the sample on a magnetic rack for0h 2m 0s (or longer if necessary) until the solution becomes clear and the beads form a pellet.

While leaving tubes on the magnet, carefully remove and discard supernatant without disturbing the beads using a P1000.

While the tube is on the magnetic rack, add 200µLof Pronex wash buffer without flushing directly onto the pellet. If 200µLis not enough to submerge the pellet, use more wash buffer.

Allow to incubate for 0h 0m 30s-0h 1m 0s .

While leaving tubes on the magnet, carefully remove and discard wash buffer without disturbing the beads using a P1000.

Keeping the tube on the magnetic stand, carefully add 200µL of Pronex Wash Buffer without flushing directly onto the pellet. If 200µL is not enough to submerge the pellet, use more wash buffer.

Allow to incubate for 0h 0m 30s-0h 1m 0s .

While leaving tubes on the magnet, carefully remove and discard wash buffer without disturbing the beads using a P1000.

Allow the sample to air dry with lids open for 0h 2m 0s-0h 5m 0swatching it until the pellet is no longer shiny.

Note

Remove the sample from the magnetic stand.

Remove the tube from the magnetic rack and add 32µL of nuclease free water. Resuspend the beads by slowly pipetting or stirring with the pipette tip.

Note

Be as gentle as possible while ensuring that pellet is resuspended.

Leave for0h 5m 0s at Room temperature to elute the DNA.

Pellet beads on magnet for 0h 1m 0s until solution becomes clear and slowly pipette DNA eluate into a new LoBind tube.

Note

Save the tube with Pronex beads in case of incomplete DNA elution, so you can repeat.

Quantify 1µL of DNA elute using the Qubit™ dsDNA HS Assay Kit or QuantiFluor® ONE dsDNA System.