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dA (Pac) CPG Column
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dA (Pac) CPG Column
CPG column for incorporation of unmodified dA at 3' end of an oligonucleotide.
Key features
Show- CPG has long-chain alkylamino succinyl linker
- Various column formats available in different synthesis scales.
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Product information
Various DNA CPG column formats are available, and compatible with a range of instruments.
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Synthesizer
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Column
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Type/Description
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Notes
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|---|---|---|---|
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MerMade 6,12
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MerMade, syringe (all scales)
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Pipette type column
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A MerMade column is also known as a Supercolumn
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MerMade 4, 48X, 96E, 192E, 192X
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MerMade, Syringe (up to 1.3 mL)
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Pipette type column
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A MerMade column is also known as a Supercolumn
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ABI 384 / 394
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Luer
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Barrel column with luer fitting at either end
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Also known as ALL-FIT or Standard
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Expedite 8909
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Luer
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Barrel column with luer fitting at either end
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Also known as ALL-FIT or Standard
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ABI3900
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MerMade
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Pipette type column
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A MerMade column is also known as a Supercolumn
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K&A H4, H8, H8SE, H2, H32, H64
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Luer
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Barrel column with luer fitting at either end
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Also known as ALL-FIT or Standard
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K&A S4CL/S8CL
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Luer
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Barrel column with luer fitting at either end
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Also known as Standard. For this instrument, we recommend the Luer (Standard) column as the ALL-FIT columns have a wider barrel.
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Dr Oligo 48
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MerMade
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Pipette type column
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A MerMade column is also known as a Supercolumn
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Dr Oligo 192XLc, 768XLc just plates
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MerMade, Syringe (up to 1.3 mL)
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Pipette type column
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A MerMade column is also known as a Supercolumn
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OligoMaker X12, 48, 192, X192, X96
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MerMade, Syringe (up to 1.3 mL)
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Pipette type column
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A MerMade column is also known as a Supercolumn
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Applicable Products
| LK2059 | Pac-dA-CE Phosphoramidite |
| LK2060 | iPr-Pac-dG-CE Phosphoramidite |
| LK2290 | Pac-dA-SynBase™ CPG 1000/110 |
| LK2292 | iPr-Pac-dG-SynBase™ CPG 1000/110 |
| LK4210 | Cap Mix A: THF/pyridine/Pac-anhydride (85:10:5) |
General
Although Ac-dC-CE Phosphoramidite (LK2034) can be deprotected under UltraMILD conditions this monomer is routinely used under standard and UltraFAST conditions. Customers should refer to the standard protocols for use of this product and the analogous CPG supports.
Physical & Dilution Data
Dilution volumes (in ml) are for 0.1M solutions in dry acetonitrile (LK4050). Adjust accordingly for other concentrations. For µmol pack sizes, products should be diluted as 100µmol/ml to achieve 0.1M, regardless of molecular weight.
Item |
Mol. Formula |
Mol. Wt. |
Unit Wt. |
250mg |
500mg |
1g |
| LK2059 | C48H54N7O8P | 887.97 | 313.21 | 2.82 | 5.63 | 11.26 |
| LK2060 | C51H60N7O9P | 946.05 | 329.21 | 2.64 | 5.29 | 10.57 |
| LK2290 | - | - | 313.21 | - | - | - |
| LK2292 | - | - | 329.21 | - | - | - |
Coupling
No changes are required from the standard synthesiser procedure. Likewise, supports are be treated as normal. If many dG residues are included in the oligonucleotide, we recommend the use of phenoxyacetic anhydride in Cap Mix A (LK4210). This removes the possibility of exchange of the iPr-Pac protecting group on the dG moiety with acetate from the standard acetic anhydride capping mix.
Deprotection & Purification
We no longer stock UltraMILD Deprotection solution (0.05M Potassium Carbonate in Methanol). This is because in our experience this reagent works best when freshly prepared just prior to use. To prepare 100ml of such a solution:
- To a suitable volumetric calibrated vessel, add 0.69g potassium carbonate (K2CO3).
- Add dry methanol to a volume of 100ml.
- Stir, protected from moisture, until dissolved leaving overnight if necessary.
Cleavage and deprotection can be carried out in 4-17h at room temperature with this solution (alternatively use AMA at room temperature for 2h; ultimately the overall protocol will be dependent on the requirements of the base-sensitive modifications being used). Cartridge purification can be incorporated into the deprotection protocol.
Typical Protocol
- Carry out the synthesis of the modified oligonucleotide.
- Open the synthesis column and transfer the support to a suitable reaction vial.
- Treat the support with 0.5ml of 0.05M potassium carbonate in anhydrous methanol for a minimum of 4h at room temperature. For oligos with a high dG content reaction overnight is recommended.
- Desalt with a G25 column.
Storage & Stability
Phosphoramidites and supports are stored refrigerated at 2 to 8°C. Phosphoramidite solutions should be used within 24h.
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