Why Yield Drops Faster Than You Expect
Even with 99% coupling efficiency, yields compound quickly. A 100-base oligo may produce less than 40% crude yield — before purification.
Improving Coupling Efficiency
- Eliminate moisture
- Optimize reagent volumes and timing
- Choose appropriate solid supports
- Maintain fresh reagents
Cleavage and Deprotection
Cleavage releases the oligo from the solid support, while deprotection removes chemical protecting groups. Conditions vary based on oligo type and modifications.
RNA oligos require an additional 2′-deprotection step, making protocol selection especially important.
Purification Options
Common methods include:
- Desalting (low purity)
- Cartridge purification
- Reverse-phase HPLC
- Anion-exchange HPLC
- PAGE for long or highly structured oligos
The right method depends on oligo length, application, and purity requirements.
Final Thought
Understanding yield, purification, and analytical methods ensures you select the correct synthesis scale and avoid costly rework.
