Coupling: No changes needed from standard method recommended by synthesizer manufacturer.
Deprotection: Treat the CPG with 10% DBU in anhydrous methanol at Room Temperature for 5 days in the dark to cleave and deprotect the oligo.
1) Prepare a solution of 10% DBU in anhydrous methanol. DBU (1,8-Diazabicyclo[5.4.0]undec-7-ene) is available from Aldrich (139009-25G) as is anhydrous methanol (322415-100ML). Note: 10% DBU in anhydrous methanol is not compatible with dmf-protecting groups.
2) Transfer the CPG to a clean, dry glass vial and add 1 mL of the 10% DBU solution.
3) Seal the vial and leave for 5 days in the dark at room temperature.
4) Transfer the contents to a micro centrifuge tube and reduce under vacuum to a small volume.
5) Add 1mL of 10mM aqueous sodium hydroxide solution and desalt or purify the oligonucleotide using standard procedures.
Storage: Refrigerated storage, maximum of 2-8°C, dry
The table below shows pack size data and, for solutions, dilutions and approximate couplings based on normal priming procedures. Please link for more detailed usage information with the various synthesizers.