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dA (Pac) CPG Column

dA (Pac) CPG Column

CPG column for incorporation of unmodified dA at 3' end of an oligonucleotide.

Key features

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  • CPG has long-chain alkylamino succinyl linker
  • Various column formats available in different synthesis scales.
Option 1: Select a Pore Size
Option 2: Select a Column Type
Option 3: Select a Scale
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Product information

Various DNA CPG column formats are available, and compatible with a range of instruments. 

Synthesizer
Column
Type/Description
Notes
MerMade 6,12
MerMade, syringe (all scales)
Pipette type column
A MerMade column is also known as a Supercolumn
MerMade 4, 48X, 96E, 192E, 192X
MerMade, Syringe (up to 1.3 mL)
Pipette type column
A MerMade column is also known as a Supercolumn
ABI 384 / 394
Luer
Barrel column with luer fitting at either end
Also known as ALL-FIT or Standard
Expedite 8909
Luer
Barrel column with luer fitting at either end
Also known as ALL-FIT or Standard
ABI3900
MerMade
Pipette type column
A MerMade column is also known as a Supercolumn
K&A H4, H8, H8SE, H2, H32, H64
Luer
Barrel column with luer fitting at either end
Also known as ALL-FIT or Standard
K&A S4CL/S8CL
Luer
  Barrel column with luer fitting at either end 
 Also known as Standard. For this instrument, we recommend the Luer (Standard) column as the ALL-FIT columns have a wider barrel.
Dr Oligo 48
MerMade
Pipette type column
A MerMade column is also known as a Supercolumn
 Dr Oligo 192XLc, 768XLc just plates 
 MerMade, Syringe (up to 1.3 mL) 
Pipette type column
A MerMade column is also known as a Supercolumn
 OligoMaker X12, 48, 192, X192, X96 
MerMade, Syringe (up to 1.3 mL)
Pipette type column
A MerMade column is also known as a Supercolumn

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:

  1. To a suitable volumetric calibrated vessel, add 0.69g potassium carbonate (K2CO3).
  2. Add dry methanol to a volume of 100ml.
  3. 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

  1. Carry out the synthesis of the modified oligonucleotide.
  2. Open the synthesis column and transfer the support to a suitable reaction vial.
  3. 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.
  4. 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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