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===Step 3. Prepare the Streptavidin Dynabeads=== #Allow the Streptavidin Dynabeads to warm to room temperature for 30 minutes prior to use. #Mix the beads thoroughly by vortexing for 1 minute. #Aliquot 100μl of beads for each capture into a single 1.5ml tube (i.e. for 1 capture use 100μl beads and for 4 captures use 400μl beads, etc.). Enough beads for 6 captures can be prepared in a single tube. #Place the tube in a DynaMag-2 device. When the liquid becomes clear (should take less than 5 minutes), remove and discard the liquid being careful to leave all of the beads in the tube. Any remaining traces of liquid will be removed with subsequent wash steps. #While the tube is in the DynaMag-2 device, add twice the initial volume of beads of Streptavidin Dynabead Binding and Wash Buffer (i.e. for 1 capture use 200μl of buffer and for 4 captures use 800μl buffer, etc.). #Remove the tube from the DynaMag-2 device and vortex for 10 seconds. #Place the tube back in the DynaMag-2 device to bind the beads. Once clear, remove and discard the liquid. #Repeat Steps 3.5 - 3.7 for a total of 2 washes. #After removing the buffer following the second wash, resuspend by vortexing the beads in 1x the original volume using the Streptavidin Dynabead Binding and Wash Buffer (i.e. for 1 capture use 100μl buffer and for 4 captures use 400μl buffer, etc.) that was prepared in Step 2.3. #Aliquot 100μl of resuspended beads into new 0.2ml tubes. #Use the DynaMag-2 device to bind the beads by holding the tube against the magnet. Remove and discard the liquid when clear. #The Streptavidin Dynabeads are now ready to bind the captured DNA. Proceed immediately to “Step 4, Bind DNA to the Streptavidin Dynabeads.”
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