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| Oligonucleotide purification
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| [ 文章来源： | 文章作者：
| 发布时间：2006-09-27|
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1. Add 400 ul of TE buffer then 400 ul 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 ul 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 ul of 5M KAc and fill tube with 95% ETOH.
10. Precipitate for at least one hour at -70¹C.
11. Spin for 15 minutes/ 4¹C/ maximum speed.
12. Pour off ETOH and dry in a speed vac as above.
13. Add 500 ul 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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Oligo - Storag