Dimethylformamide → (2S)-2,6-bis((((tert-butoxy)carbonyl)amino))hexanoic acid

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Reaction Steps

Di-(t-Butoxycarbonylamino)-L-lysine (0.11 g, 20 μmole), benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP) (0.10 g, 20 μmole), Hydroxybenzotriazole (HOBt) (0.03 g, 20 μmole) and N-methylmorpholine (NMM) (0.04 mL, 40 μmole) were dissolved in 1 mL dimethylformamide (DMF) and stirred for 1 minute. Then the mixture was contacted with beads to which the amino linker oligonucleotides (1 μmole) prepared in Example 1 were bound, and stirred for 1 hour. After being washed with dimethylformamide and dichloromethane, each three times, the bead was placed in 25% TFA/CH2Cl2 and stirred for 30 minutes. The bead was washed with dimethylformamide and dichloromethane, each three times. Biotin (0.05 g, 20 μmole), benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP) (0.10 g, 20 μmole), Hydroxybenzotriazole (HOBt) (0.03 g, 20 μmole) and NMM (0.04 mL, 40 μmole) were dissolved in 1 mL dimethylformamide (DMF) and were stirred for 1 minute. Then, the mixture was contacted with beads to which the amino linker oligonucleotides (1 μmole) were bound. The beads were stirred for 1 hour, and washed with dimethylformamide and dichloromethane, each three times. To the beads, 2 mL of saturated NH4OH was added and placed at 60° C. for 8 hours. The solid part was isolated out from the centrifuge, and the liquid part was collected and dried under vacuum. The obtained biotin-labeled oligonucleotides were purified by RP-HPLC and freeze-dried, to produce the pure entitle compound in solid phase.

Provenance & Evidence