Oligonucleotides can be prepared on the DNA synthesizer either trityl-on or
trityl-off
(trityl = DMT) by making the selection at the beginning of the synthesis.
Virtually all applications require DNA to be trityl-off, or detritylated, so
that they have a free 5' hydroxyl.
However, the trityl-on option is selected when purifying using an OPC
cartridge or certain types of HPLC. After purification, trityl-on
oligonucleotides must be detritylated manually. The 5' dimethoxytrityl (DMT) is
readily cleaved by limited exposure to mild acidic conditions to minimize
depurination.
Materials:
Procedure
- Dissolve the dried, deprotected oligo in 1 ml 80% acetic acid. Mix by
vortexing.
- Incubate for 1 hour at room temperature.
- Dry the detritylated oligo using vacuum centrifugation.
- A reverse-phase desalting procedure may then be used to remove the free
DMT from the oligo solution.
Reference
“Evaluating and Isolating Synthetic Oligonucleotides-The Complete Guide” Applied
BioSystems, Inc., 1992, p A-7.