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Bz-S-C6-dT CE-Phosphoramidite
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Bz-S-C6-dT CE-Phosphoramidite
Phosphoramidite for the incorporation of a thiol function internally within an oligonucleotide.
Key features
Show- Incorporates a thiol reactive functional group for conjugation internally within an oligonucleotide.
- Avoids need for conversion of e.g. amino functionality to thiol.
- Can be directly conjugated to maleimides and haloacetamides.
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Product information
Incorporation of a thiol reactive functional group at specific sites within an oligonucleotide allows for subsequent post-synthesis conjugation of the oligo with a number of different moieties such as fluorescent markers and biotin, depending on the application. Such labels need to be reactive towards the incorporated functional group: for example, thiols will react with iodoacetate and maleimide derivatives to form thioether linkages.
Thiol modification has classically been limited to the 5’ or 3’ end of the oligonucleotide, however it is now possible using this thiol-dT modification that can be incorporated within an oligonucleotide and reacts directly with maleimides and haloacetamides.
Applicable Products
| LK2071 | Amino-Modifier-C6-dA-CE Phosphoramidite |
| LK2135 | Amino-Modifier-C6-dT-CE Phosphoramidite |
| LK2141 | Amino-Modifier-C6-dC-CE Phosphoramidite |
| LK2149 | Amino-Modifier-C2-dT-CE Phosphoramidite |
| LK2191 | Bz-S-dT-CE Phosphoramidite |
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 |
| LK2071 | C55H65F3N9O8P | 1068.14 | 427.40 | 2.34 | 4.68 | 9.36 |
| LK2135 | C50H62N6O10F3P | 995.05 | 458.41 | 2.51 | 5.02 | 10.05 |
| LK2141 | C53H68F3N8O9P | 1049.14 | 457.42 | 2.38 | 4.77 | 9.53 |
| LK2149 | C46H54N6O10F3P | 938.94 | 402.30 | 2.66 | 5.33 | 10.65 |
| LK2191 | C58H71N6O11PS | 1091.27 | 546.53 | 2.29 | 4.58 | 9.16 |
Amino Modifiers (LK2071, LK2141, LK2149 & LK2135)
Coupling
No changes are required from the standard method recommended by the synthesiser manufacturer. Coupling is as per standard nucleoside amidites.
Cleavage & Deoprotection
The trifluoroacetyl (TFA) protecting group on the primary amine is removed during standard ammonium hydroxide solution deprotection. However, a minor side reaction during deprotection can lead to irreversibly capping 2-5% of the amine. This could be significant if multiple additions of the modifier are made.
To prevent the reaction, synthesise using acetyl-protected C and, prior to cleavage and deprotection, wash the column with 10-20% DEA/acetonitrile. Deprotect in AMA at 65°C for 15min.
Thiol Modifier (2191)
Coupling
BTT or DCI can be used as activators. Do not use ETT. A coupling time of 10min (600s) is recommended. To facilitate AMA deprotection, dmf-G and Ac-C should be used.
Cleavage, Deprotection & Purification
Before carrying out the cleavage and deprotection, wash with a 20% DEA/MeCN solution, then:
- Cleave and deprotect using 100mM TCEP in AMA at room temperature for 2h.
- Desalt using a G25 column.
- Purify if required.
- Dry the oligo.
- Add 100μl of 87mM TCEP in water.
- Leave for 1h at room temperature.
- Pass through a G25 column pre-equilibrated with the conjugation buffer.
Conjugation
To conjugate, add the appropriate maleimide, acetamide or equivalent to the above solution and react as appropriate.
Once the reaction is complete, pass through a G25 column pre-equilibrated with 0.1M TEAA or, if conjugation is on a solid surface, thoroughly with water.
All products
Storage & Stability
Store in a freezer below -10°C. Diluted samples must be freshly prepared for use and used within 24h.
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