SuperPure™ Purification Column (<= 50 nmol scale synthesis)
货号：SP-1000-1,SP-1000-96
规格：1,plate/96
价格：24.656,2110.5
产品类型：核酸合成
品牌：LGC Biosearch Technologies
| Oligonucleotides (whether synthetic or natural) may be purified by a number of techniques, including
polyacrylamide gel electrophoresis and high performance liquid chromatography (either by ion exchange or reverse-phase methods). Synthetic DNA can be conveniently de-salted and purified by reverse-phase low-pressure separation on BTI's SuperPure (SP-1000) columns. BTI's SuperPure Columns are intended for reverse-phase purification of 5'-DMT oligonucleotides followed by on-column detritylation. The method relies on strong binding between the 5'-DMT protected product and a hydrophobic optimized polymer packing (polystyrene), thus allowing separation from truncated sequences which, due to a lack of DMT group, do not bind. Treatment with acid removes the DMT group from the product, the desired product (free from organic residues) is eluted with 20% acetonitrile. The method is rapid, efficient and permits many samples to be purified simultaneously. The SuperPure column has capacity to handle crude, cleaved DNA from a 50 nmol scale synthesis. The purified yield will depend, in each case, on sequence length and the quality of the crude sample. |
| 产品用途： |
| 1. Program DNA synthesizer to leave terminal 5'-DMT group attached;
2. Cleave and deprotect product with conc. aqueous ammonia (5 hours at 60 °C for standard deprotection procedure);
3. Evaporate cleavage solution to dryness;
4. Re-suspend in 0.2 ml 0.1N TEAA or H2O;
5. Pre-equilibrate column by passing 2 column volumes (CVs) of acetonitrile through the column;
6. Repeat with 3 CVs 1N TEAA (triethylamine acetate ;
7. Take the oligo solution of step 4 and apply slowly to the column while collecting effluent in a separate
micro-centrifuge tube. Reload effluent and pass once more through column (optional);
8. Pass 3 CVs of 2.8% ammonia solution slowly through the column;
9. Wash by slow passage of 2 CVs water;
10. Pass 2 CVs of 2.5% TFA through the column, wait 5 minutes;
11. Pass 3 or more CVs of water through column. Check to see pH is neutral.
12. Slowly pass 2 CVs 20% acetonitrile through the column, collecting effluent containing the desired
purified product in a labeled microcentrifuge tube;.
13. Speedvac ™ the solution to dryness, analyze and use as desired. |
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