DNA Extraction of Cesium Chloride-Purified Viruses using Wizard Prep Columns
Prepare 2 labeled 1.5ml microfuge tubes per sample and make sure resin is at room temperature.
Shake resin vigorously to resuspend particles.
Mix 1ml DNA Purification Resin with 0.5ml CsCl sample.
Attach minicolumn to bottom of 3ml or 5ml sterile syringe that has had plunger removed.
Add resin with sample to the syringe and push through the solution.
Remove minicolumn from the syringe and pull out plunger.
Reattach minicolumn to the syringe and 2ml of 80% isopropanol to the syringe.
Using the plunger push through the isopropanol to wash the resin.
Remove minicolumn from syringe and place in a sterile 1.5ml centrifuge tube.
Centrifuge 10,000 g for 2min to remove any residual liquid.
Place minicolumn in new sterile 1.5ml centrifuge tube.
Add 100μl 80°C TE buffer to top of minicolumn.
Place tube lid over top of column and vortex gently for 10 seconds.
Wait 1 minute.
Centrifuge at 10,000 g for 30 sec to elute DNA.
Quantify DNA by Quant-iT PicoGreen dsDNA assay kit.
