Please reduce the thiol before using the oligo in your application:
1. Add 200 µl 10mM TCEP (solve 2,9mg TCEP-HCL in 1ml 1xTE buffer, pH 8.0) to your oligo
2. Shake it for 60 min at room temperature
3. Precipitate the oligo:
- Add 150 µl 3 M Na-Acetate [24,6 g Na-Acetate + 0,21 g Mg-Acetate in 100 ml dest. H2O]
- Fill tube with ethanol p.a.
- Shake gently
- Incubate 20 min at –20°C
4. Spin for 5 min at 13000 rpm, discard supernatant
5. Dry pellet at room temperature
Please note:
After the treatment the molecular wight of the thiol-modiifcation will change:
[ThiolC3]: after reduction: 90.14 (C3H6OS); before: 180.28 (C6H12O2S2)
[ThiolC6]: after reduction: 132.22 (C6H12OS); before: 264.44 (C12H24O2S2)