| Reagent |
Quantity (for 500 μL) |
Final concentration |
| rATP (not dATP) (100 mM) |
50 μL |
10 mM |
| rCTP (not dCTP) (100 mM) |
50 μL |
10 mM |
| rGTP (not dGTP) (100 mM) |
50 μL |
10 mM |
| rUTP (65 mM) |
50 μL |
6.5 mM |
| Digoxigenin-11-UTP (or fluorescein-12-UTP or biotin-16-UTP) (35 mM) |
50 μL |
3.5 mM |
|
| The labeling mix may be purchased as a 10X digoxigenin-rUTP/rNTP mix (Roche), or it can be prepared according to the recipe.
For detection, use anti-digoxigenin-alkaline-phosphatase (AP) antibody (Roche), anti-fluorescein-AP antibody (Roche), or strepavidin-AP
(Roche), respectively. Store in aliquots at −20°C.
|
