Use of Bz-S-C6-dT-CE Phosphoramidite (2191) in Oligonucleotide Synthesis
Dilution
Dilute in anhydrous acetonitrile to a concentration of 0.1M (250mg pack in 2.29ml, 100µmol pack (109mg) in 1ml).
Coupling
BTT or DCI can be used as activators. Do not use ETT. A coupling time of 10min (600s) is recommended.
Oxidation
No changes are required from standard methods.
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.
Storage & Stability
Store in a freezer below -10°C. Diluted samples should be freshly prepared for use and used within 24h.