Applicable Products
9003 | TOP-DNA Tubes (150mg) |
9004 | TOP-RNA Tubes (100mg) |
9005 | TOP-DNA Jnr, Bond Elut (150mg) |
9006 | TOP Tubes (50mg) |
Solution Preparation details
100mg/mL NaCl
- Weigh 10g of sodium chloride into a 100mL volumetric flask.
- Add water to give a total volume of 100mL.
- Mix thoroughly.
2% TFA solution
NOTE this solution needs to be made fresh on the day of use.
- Measure 2mL of TFA and add to a 100mL bottle.
- Add 98mL of water.
MeCN/water
- Measure 500mL of MeCN and add to a 1L bottle.
- Measure 500mL of water and add to the MeCN.
- Mix thoroughly.
2M Tris HCl(aq)
- Weigh out 31.53g into a 100mL volumetric flask.
- Add water to give a final volume of 100mL.
- Mix thoroughly.
1M ammonium bicarbonate / 30% MeCN
- Weigh out 7.91g into a 100mL volumetric flask.
- Add water to give a final volume of 100mL.
- Mix thoroughly.
TOP-DNA Purification
Do not remove the deprotection solution or desalt the oligo. The deprotection solution needs to be present.
- Add 1mL of aqueous sodium chloride solution (100mg/mL) to each oligo.
- Place one column for each oligo onto the vacuum manifold.
- Turn the vacuum pump on.
- Adjust the pressure to 7.0in (178mm) Hg using the vacuum control valve.
- Add 0.5mL of acetonitrile to the cartridge.
- Immediately then add 1mL of 2MTEAA to condition the medium.
- Add the oligo solution to the cartridge in 1mL aliquots.
- Add 1mL of NaCl(aq) solution (100mg/mL) to the column.
- Add 1mL of NaCl(aq) solution (100mg/mL) to the column.
- Add 1mL of 2% TFA/water solution to the column.
- Add 1mL of 2% TFA/water solution to the column.
- Add 1mL of water to the column.
- Add 1mL of water to the column.
- Release the vacuum.
- Place an appropriately labelled tube beneath each of the column.
- Add 1mL of MeCN/water 1:1 to elute the DNA oligo under vacuum.
TOP-DNA Jnr (Bond Elut®) Purification
Do not remove the deprotection solution or desalt the oligo. The deprotection solution needs to be present.
- Add 1mL of aqueous sodium chloride solution (100mg/mL) to each oligo.
- Appropriately label one cartridge for each oligo to be purified.
- Pass 0.5mL of acetonitrile slowly through the cartridge.
- Immediately then pass 1mL of 2MTEAA to condition the medium.
- Add the oligo solution to the cartridge in 1mL aliquots by slowly passing through with a syringe.
- Slowly pass 1mL of NaCl(aq) solution (100mg/mL) through the cartridge.
- Slowly pass 1mL of NaCl(aq) solution (100mg/mL) through the cartridge.
- Slowly pass 1mL of 2% TFA/water solution through the cartridge.
- Slowly pass 1mL of 2% TFA/water solution through the cartridge.
- Slowly pass 1mL of water through the cartridge.
- Slowly pass 1mL of water through the cartridge.
- Place an appropriately labelled tube beneath each of the cartridges.
- Slowly pass 1mL of MeCN/water 1:1 to elute the DNA oligo collecting into the labelled tube.
TOP-RNA Purification (with TBDMS RNA Chemistry)
Do not remove the deprotection solution or desalt the oligo. The deprotection solution needs to be present. Use RNase free solutions for this procedure.
- Add 1.75mL of aqueous 2M TrisHCl solution to each oligo.
- Place one column for each oligo onto the vacuum manifold.
- Turn the vacuum pump on.
- Adjust the pressure to 7.0in (178mm) Hg using the vacuum control valve.
- Add 0.5mL of acetonitrile to the cartridge.
- Immediately then add 1mL of 2M TEAA to condition the medium.
- Add the oligo solution to the cartridge in 1mL aliquots.
- Add 1mL of MeCN/2MTEAA, pH7.0 solution (1:9, v:v) to the column.
- Add 1mL of water to the column.
- Add 1mL of 2% TFA/water solution to the column.
- Add 1mL of 2% TFA/water solution to the column.
- Add 1mL of water to the column.
- Add 1mL of water to the column.
- Release the vacuum.
- Place an appropriately labelled tube beneath each of the column.
- Add 1mL of 1M ammonium bicarbonate(aq)/30% MeCN solution to elute the RNA oligo under vacuum.
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