#D4014 from Zymo Research
1) In a 1.5 mL microfuge tube, add 2 volumes of DNA binding buffer for each one volume of DNA to be cleaned. Mix by vortexing.
For example, if you had 100 µL of DNA, add 200 µL of DNA binding buffer.
2) Transfer DNA solution to Zymo-Spin Column inserted into a 2 mL collection tube.
3) Spin at full speed for 30 seconds. Discard flow-through since the DNA is now bound to the column. The maximum volume of a spin column is 800 µL, but you can load it more than one time with the same DNA solution.
4) Add 200 µL of Wash Buffer to the column. Sping at full speed for 30 seconds. Discard flow-through.
5) Add another 200 µL of Wash Buffer to the column. Sping at full speed for 30 seconds. Discard flow-through.
6) Add 10 µL of elution buffer (10 mM Tris, pH 8.0, 1 mM EDTA) to the column. Make sure the elution buffer hits the center of the white column matrix. Transfer each column to an appropriately labeled 1.5 mL microfuge tube. Spin full speed for 30 seconds to elute the DNA. Dispose of the column since the DNA is now in your microfuge tube.
Yield of bigger fragments (> 6 kb) can be improved by letting the elution buffer stand for 1 minute before spinning. Fragments > 10 kb can be eluted more efficiently with warm (65° C) TE.
7) OD the purified DNA with the Nanodrop. Take 0.5 µL of the DNA, mix with 1.5 µL of elution buffer, and OD 2 µL with TE as your blank. If the DNA is too concentrated (i.e., OD260 > 1.5), then dilute the remaining 1 µL of DNA so the new OD260 will be between 0.05 and 1.5 on the Nanodrop.
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