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        Oligonucleotide purification

        互联网

        897

        1. Add 400 μl of TE buffer then 400 μl of 1-butanol to the oligonucleotide glass vial.

        2. Vortex well, then spin down in tabletop centrifuge for 10 minutes at ~2,000. rpm¹s.

        3. Remove top, butanol layer with a sterile pipette tip and discard.

        4. Add another 400 μl of 1-butanol.

        5. Vortex well, then spin down as above in tabletop centrifuge.

        6. Remove top, butanol layer and discard.

        7. Transfer aqueous phase into a new 1.5 ml tube.

        8. Dry in a speed vac for about 5 minutes to remove all of the butanol.

        9. Add 50 μl of 5M KAc and fill tube with 95% ETOH.

        10. Precipitate for at least one hour at -70℃.

        11. Spin for 15 minutes/ 4℃/ maximum speed.

        12. Pour off ETOH and dry in a speed vac as above.

        13. Add 500 μl of TE buffer to your oligo. in the 1.5 ml tube and mix well.

        14. Dilute 1:10 and/or 1:100 and read on spectrophotometer to determine concentration.

        15. Follow product specification sheet to determine volume needed to make a 20 um solution.

         

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