Buffer NTINucleoSpin\u00ae Gel and PCR Clean-up Column NucleoSpin\u00ae Gel and PCR Clean-up Collection Tube (2ml)Buffer NT3 Buffer NE \/ nuclease-free waterSolubilize gel sliceFor each 100 mg of agarose gel < 2 % add 200 \u03bcL Buffer NTI. For gels containing > 2 % agarose, double the volume of Buffer NTI. Incubate sample for 5\u201310 min at 50 \u00b0C. Vortex the sample briefly every 2\u20133 min until the gel slice is completely dissolved! Bind DNAPlace a NucleoSpin\u00ae Gel and PCR Clean-up Column into a Collection Tube (2 mL) and load up to 700 \u03bcL sample. Centrifuge for 30 s at 11,000 x g. Discard flow-through and place the column back into the collection tube. Load remaining sample if necessary and repeat the centrifugation step. Wash silica membrane Add 700 \u03bcL Buffer NT3 to the NucleoSpin\u00ae Gel and PCR Clean-up Column. Centrifuge for 30 s at 11,000 x g. Discard flow-through and place the column back into the collection tube. Dry silica membrane Centrifuge for 1 min at 11,000 x g to remove Buffer NT3 completely. Make sure the spin column does not come in contact with the flow-through while removing it from the centrifuge and the collection tube. Note: Residual ethanol from Buffer NT3 might inhibit enzymatic reactions. Total removal of ethanol can be achieved by incubating the columns for 2\u20135 min at 70 \u00b0C prior to elution. Elute DNAPlace the NucleoSpin\u00ae Gel and PCR Clean-up Column into a new 1.5 mL microcentrifuge tube (not provided). Add 15\u201330 \u03bcL Buffer NE and incubate at room temperature (18\u201325 \u00b0C) for 1 min. Centrifuge for 1 min at 11,000 x g.