Paul.xu at 20:11, 15 December 2018

2018-12-15T20:11:56Z

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Prepare 20 mg/ml   (20 ug/ul) Glycogen solution in Nuclease-free water.

Weigh 100 mg, Nuclease-free water add 5 ml H<sub>2</sub>O, transfer 0.5 ml into microcentrifuge tubes, store at -20 <sup>o</sup>C

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.
[[Category:Protocols]]

Precipitation of DNA using Glyogen - Revision history

Paul.xu at 20:11, 15 December 2018