Ligation of a DNA or RNA oligo to single stranded RNA using T4 RNA Ligase 1
Component | 20 μl reaction |
ssRNA acceptor* | 1-20 pmol |
RNA or DNA oligo donor** | 5-40 pmol |
10X T4 RNA ligase buffer | 2 ul (1X) |
10 mM ATP | 2 ul (1 mM) |
50% PEG8000 | 5-10 ul (final 15-25%) |
Rnase inhibitor (40 u/ul) | 1 ul (2 u/ul) |
T4 RNA ligase 1 Enzyme | 1 ul (10 units) |
Nuclease-free Water | To 20 μl |
Incubation Time & Temperature | 2 hour at 25°C or 16 hour at 16°C |
Enzyme should be added at the last step.