3'-Amino Modifier C7 CPG Column
3'-Amino Modifier C7 CPG Column
3'-Amino Modifier C7 CPG Column, 1000 Å, 200 nmol (Pack of 4), ALL-FIT
CPG column for the incorporation of an amino function at the 3' end of an oligonucleotide.
Key features
Show- Incorporates a primary amine at the 3 end of an oligonucleotide, for subsequent conjugation.
- Most commonly used product for introducing a 3'-amino functionality.
- Fmoc protection is quite stable to oligo synthesis conditions, however careful handling is required to prevent loss of Fmoc leading to capping of the free amine with acetic anhydride and hence loss of functionality.
- Fmoc can be removed selectively without cleavage from the support allowing solid-phase conjugation of the desired label.
- CPG has long-chain alkylamino succinyl linker.
Usually shipped in 3-5 days, Packed To Order
Product information
Incorporation of a primary amine reactive functional group at specific sites within an oligonucleotide allows for subsequent post-synthesis conjugation of the oligo with a number of different affinity, reporter or protein labels, depending on the application. Such labels need to be reactive towards the incorporated functional group: for example, NHS esters or isothiocyanates will react with primary amines. This approach is often necessary where the desired label or tag is either not available as a phosphoramidite, or is sensitive or unstable to the conditions of oligonucleotide synthesis or deprotection. A common example is the attachment of a rhodamine dye using the TAMRA NHS ester. Functionally-derivatised oligos can also be covalently attached to surfaces such as glass slides or gold microspheres for use in various microarray or nanoelectronic applications.
The most commonly used product for introducing a 3'-amino functionality is Fmoc-protected 3'-amino C7 CPG. Use of an Fmoc-protected amine has both advantages and disadvantages. This is quite stable to oligo synthesis conditions however, if not handled correctly, some loss of Fmoc may occur. This leads to capping of the free amine with acetic anhydride and hence loss of functionality. The main advantage of Fmoc is that it can be removed selectively without cleavage from the support allowing solid-phase conjugation of the desired label. This can be done prior to, or subsequent to, oligonucleotide synthesis.
Alternatives to Fmoc protection have been investigated. Phthalimide (PT) chemistry has been used in the development of 3'-PT-amino-C6 CPG (1), where the nitrogen that will ultimately provide the 3'-amino function is part of the PT group attached to the support through an imide group attached to the aromatic ring. This linkage is stable to all conditions of oligo synthesis and the resulting amino functionality does not add any additional chiral centres/diastereomers to the oligo. A completely analogous C3 product is also available.
Additional products are available for introducing 3'-amino functionality without blocking the terminus from any desired enzymatic activity. There are the C6 dC and dT analogues, plus the mdC TEG product confers additional spacing and hydrophilicity. These products all have base-labile trifluoroacetyl (TFA) protection on the amine, which is removed during cleavage and deprotection.
Ref:
- An improved CPG support for the synthesis of 3'-amine-tailed oligonucleotides, C.R. Petrie, M.W. Reed, A.D. Adams and R.B. Meyer, Jr., Bioconjugate Chem., 3, 85-87, 1992.
Synthesizer
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Column
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Type/Description
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Notes
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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)
|
Pipette type column
|
A MerMade column is also known as a Supercolumn
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ABI 384 / 394
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Luer
|
Barrel column with luer fitting at either end
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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
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Luer
|
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)
|
Pipette type column
|
A MerMade column is also known as a Supercolumn
|
Applicable Products
LK2350 | 3'-Amino-Modifier-C7 CPG 1000 |
LK2365 | 3'-PT-Amino-Modifier-C6 CPG |
LK2367 | 3'-Amino-Modifier-C6-dT CPG |
LK2369 | 3'-Amino-Modifier-C6-dC CPG |
LK2371 | 3'-PT-Amino-Modifier-C3 CPG |
Physical Data
Item |
Unit Wt. |
LK2350 | 209.18 |
LK2365 | 179.15 |
LK2367 | 359.24 |
LK2369 | 457.42 |
LK2371 | 137.07 |
Coupling
All supports are used as per unmodified nucleoside supports. However, non-nucleosidic modifications are slow to detritylate and require an initial detritylation prior to use in synthesis. In this case it is important not to use a cycle with an initial capping step. AMA deprotection will require dmf-G and Ac-C to be used.
Cleavage & Deprotection
LK2350—Cleavage of the oligonucleotide requires 2h at room temperature with ammonium hydroxide or AMA. It is important to wash the column with 10-20% diisopropylamine in acetonitrile or 10-20% DEA/acetonitrile to prevent blocking with acrylonitrile during deprotection. Alternatively, 20% piperidine in DMF can be used, rather than DEA. In fact this will simultaneously remove Fmoc and cyanoethyl protection, plus remove the vinylfluorene by-product from the Fmoc deprotection reaction.
For the PT-Amino CPGs (LK2371 & LK2365)—Cleavage of the oligonucleotide requires overnight at 55°C with ammonium hydroxide. This treatment will complete the deprotection of the nucleobases for all protection types. It is important to wash the column with 10-20% diisopropylamine in acetonitrile or 10-20% DEA/acetonitrile to prevent blocking with acrylonitrile during deprotection.
LK2369 & LK2367—Use deprotection conditions as required by the nucleobases. It is important to wash the column with 10-20% diisopropylamine in acetonitrile or 10-20% DEA/acetonitrile to prevent blocking with acrylonitrile during deprotection.
Storage & Stability
The supports are stored in a freezer at –10 to –30°C.
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