How to Recycle QIAquick Purple Spin Columns

Adapted from: Siddappa et al., 2007, Biotechniques

1) When you are finished with the gel purification from Qiagen, place the used columns into the paired, used collection tubes.

2) Fill used column with 700 mL of 1 M HCl.

3) Store with cap closed in an airtight container for at least 24 hours (up to 30 days).

4) Once you have a bunch of columns and collection tubes in the 1 M HCl, to prepare a column for use:

• Thoroughly rinse the columns and collection tubes in a large beaker of dH₂O.

• Shake/ wipe off excess water. Place each column into a collection tube. Fill each column with 700 mL sterile dH₂O. Spin at maximum speed in a tabletop centrifuge for 1 minute. Discard the water.

• Repeat the previous step a second time.

• Equilibrate the column by filling it with 700 µL Buffer QBT. Spin at maximum speed in a tabletop centrifuge for 1 minute.

• Discard the buffer. Spin again for 1 minute to remove excess QBT buffer from the column.

• Put clean columns into a plastic bag. Rinse the collection tubes in a large beaker of dH₂O.

• Pour off water and allow tubes to air dry. Place into a plastic bag. The columns are now ready for use.

Note: After 9 rounds of regeneration, columns have shown optimal performance in a series of assays (Siddappa et al. 2007).

1M HCl

FW HCl = 36.47; 37% HCl = 370 g/L » 10 M

Note: for HCl, 1 Molar = 1 Normal

Qiagen Buffer QBT (Equilibration Buffer)

[750 mM NaCl, 50 mM MOPS (pH 7.0),15% ethanol, 0.15% Triton X-100]

Dissolve 43.83g NaCl, 10.46g MOPS (free acid) in 800 mL dH₂O. Adjust pH to 7.0. Add 150 mL 100% ethanol and 15 mL 10% triton X-100 solution. Adjust volume to 1 liter.

Store at room temperature.