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dG (iPr-Pac) CPG

dG (iPr-Pac) CPG

CPG for incorporation of unmodified dG at 3' end of an oligonucleotide.
  • CPG has long-chain alkylamino succinyl linker
  • Available in smaller pack sizes suitable for research use
Option 1: Select a Pore Size
Option 2: Select a Functional Loading
Option 3: Select a Size
TBD
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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
  • Suitable for high yield applications such as therapeutic oligos
  • 500 Å CPG can load up to ~100 μmol/g
1000 Å > 20mers
  • Suitable for highly modified oligonucleotides
  • modifier CPGs are functionalised onto this pore size as standard
2000 Å > 80mers
  • Can be used for CRISPR applications
  • Retains higher loading (yield) possibilities of lower pore sizes
3000 Å > 80mers
  • Can be used for CRISPR applications
  • Performs well for very long sequences > 120mers

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:

  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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