DNA Spin Columns
Regular Spin Columns
1000 rpm Ready for 25 µl -
50 µl DNA sample
· Swell 5 g Sephadex G-50 (fine) in 100 ml TE and autoclave to sterilize.
· Exclusion limit for Sephadex G-50 is 20 bp/20 nt.
· Columns work best if the sample loaded is <5% of column volume.
· Columns can be stored at room temperature for a day.
· Do not use columns that contain cracks or bubbles.
1. Remove tip of Bio-Rad column. Place column in 13 X 100 mm glass tube in rack. Fill column with
Sephadex G-50 in TE buffer. Use a sterile Pasteur pipette. Fill column from bottom to top. Tap
column to start flow and allow Sephadex to settle. Tap to coax any bubbles to the top.
2. Place column and glass tube in 50 ml plastic centrifuge tube. Spin 10 minutes at 1000 rpm
(~250 g) in IEC centrifuge. After spin, the column volume should be about 1 ml.
IEC r = ~230 mm rpm RCF rpm RCF