Precipitation of DNA using Glyogen
Prepare 20 mg/ml (20 ug/ul) Glycogen solution in Nuclease-free water.
Weigh 100 mg, Nuclease-free water add 5 ml H2O, transfer 0.5 ml into microcentrifuge tubes, store at -20 oC
1. Add 1/10 volume of 3M sodium acetate, pH 5.2 to the nucleic acid in solution (e.g., add 2.5 ul to 25 ul PCR reation)
Note: 1/10 volume of 2M sodium chloride or 5M ammonium acetate may be substituted.
2. Add Glycogen Solution, 20 mg/ml to final concentration of 0.05 ~ 1 µg/µl.
Note: For oligonucleotides and small DNA fragment, use a final concentration of 1 µg/µl.
For DNA or RNA, use a final concentration of 0.05-1 µg/µl of Glycogen Solution.
For 25 ul PCR reaction, add 1 ul of 20 mg/ml Glycogen solution (add 20 ug)
For recovery of DNA from agarose gel, add 5 ul of 20 ug/ul Glycogen stock solution (100ug)
3. Add 1 volume of isopropanol to the solution. Mix gently.
Note: 2.5 volumes of ethanol may be substituted. Note: Use ethanol for <200 bp fragments.
4. Incubate for 1 hour at -20°C or for 30 minutes at -70°C.
Note: Longer incubation times at lower temperatures may result in better recovery of nucleic acids.
5. Centrifuge for 15-30 minutes at 12,000 x g at 4°C. Discard the supernatant without disturbing the pellet.
6. Rinse the pellet with cold 70% ethanol.
7. Centrifuge for 5-15 minutes at 12,000 x g at 4°C. Discard the supernatant carefully to avoid disturbing the pellet.
8. Air dry the pellet for 5-10 minutes, being careful to not over-dry, which may render the pellet more difficult to dissolve.
Note: Isopropanol may require longer drying time than ethanol.
9. Dissolve the nucleic acid pellet in nuclease-free water, EB, or TE Buffer, pH 8.