dG (iPr-Pac) CPG
dG (iPr-Pac) CPG
Key features
Show- CPG has long-chain alkylamino succinyl linker
- Available in smaller pack sizes suitable for research use
Product information
Controlled Pore Glass (CPG) has been widely used as a solid support for oligo synthesis for several decades. LGC, Biosearch Technologies’ has perfected CPG manufacture for maximum oligo purity and yield. Our advanced CPG production techniques improve control of particle size and shape, pore size, pore volume, and specific surface area. These physical parameters influence solution exchange behaviour, ligand loading and distribution, and reaction kinetics to increase the efficiency, purity, and reproducibility of syntheses.
We have developed proprietary chemical attachment procedures to further optimise ligand distributions, providing increased accessibility to synthesis reagents and washing solutions and facilitating even better oligo yields and purities. Furthermore, process refinements and proprietary assays have been developed to minimise the troublesome “N-1” impurity levels in an oligo synthesis.
LGC, Biosearch Technologies’ Prime CPG is considered to be the gold-standard solid support used in all sectors of the market. Our collaborative process has resulted in solid supports that are optimised for the synthesis of the latest therapeutic oligo classes including LNA, delivery enhancing lipid ligands, siRNA and Spiegelmers.
Our CPG solid supports are available in a variety of pore sizes and functionalised nucleoside loadings. Which pore size and loading required is dependent on the length, complexity and application of the oligo.
In general, large scale oligo synthesis for therapeutic applications requires high loaded 500-600 Å and small to medium scale synthesis for diagnostic or research use require higher pores sizes.
Pore size | Optimal oligo length | Capabilities |
---|---|---|
500 Å and 600 Å | ≤ 30mers |
|
1000 Å | > 20mers |
|
2000 Å | > 80mers |
|
3000 Å | > 80mers |
|
Aside from Prime CPG, many research-use CPG products are also available which originated from the legacy portfolios, including SynBase™ CPGs. These CPGs are manufactured from the same glass as the Prime products, using similar processes, but generally offered in smaller pack sizes for research purposes.
SynBase loading options
Product | Average Pore Size (Å) | Nominal Particle Size (µm) | Nucleoside Loading (µmol/g) |
---|---|---|---|
SynBase™ CPG 500/50 H | 500 | 50 | 80-130 |
SynBase™ CPG 500/110 S | 500 | 110 | 35-50 |
SynBase™ CPG 500/110 H | 500 | 110 | 60-100 |
SynBase™ CPG 1000/110 | 1000 | 110 | 25-40 |
SynBase™ CPG 2000/110 | 2000 | 110 | 15-35 |
SynBase™ CPG 3000/110 | 3000 | 110 | 10-25 |
Product names are appended with either an ‘S’ or ‘H’ respectively where there is a choice
A number of linker variants are used in our DNA CPG products, although a long-chain alkyl amino succinyl linker (CNA) is the most common.
Linker variants | Benefits |
---|---|
Aminopropyl (AMP)-succinyl combination | Allows for smaller pore sizes which is ideal for research use |
glycolate linker | Cleaves under mild deprotection conditions |
Q-linker (hydroquinone-0,0'-diacetic acid group) | Quick cleavage (2 mins at room temperature) which is ideal for base sensitive oligonucleotides of dye labelled oligos |
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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