This is a cached page for the URL ( To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
DNA Purification - Removal of g-32P-ATP from Oligonucleotides

Jeff Lawrence (P20)

1. Prepare Whatman DE-52 according to manufacturers specifications. Equilibrate at store with 0.02% Sodium azide.

2. Plug a 1 mL (blue) pipet tip with Siliconized glass wool. Add 500 ml of 50:50 DE-52 slurry. Pack into a 250 mL column by gently forcing the fluid through with a P-1000 pipetter.

3. Rinse column twice with 1 mL TE, forcing through with a P-1000.

4. Add 200 ml TE to Kinased Oligo. Apply the solution to the column and gently force though with the P-1000.

5. Elute nucleotides by washing 4 times with 1 mL 0.2 M NaCL in TE. Monitor counts removed. Most residual ATP should be removed after the first 2 rinses.

6. Elute the labeled oligonucleotide with 500 ml 1 M NaCl in TE. Repeat 3 times, collecting each rinse separately. The first rinse should contain most of the oligonucleotide.

7. Up to 10% of the counts may remain on the column.