|
The optimized labelling protocol |
|
| PCR mix |
|
|
|
|
| Reagent |
Volume |
|
|
|
| Mix of DOP1,2,3 primers (15 μM) |
5 μl |
| 10× PCR Buffer w/o MgCl2 |
5 μl |
| 50 mM MgCl2 |
5 μl |
| dATP (10 mM) |
1 μl |
| dGTP (10 mM) |
1 μl |
| dCTP (10 mM) |
1 μl |
| dTTP (10 mM) |
0.7 μl |
| x- dUTP (1 mM) * |
1 μl |
| H2O |
27.8 μl |
| Platinum Taq DNA Polymerase |
0.5 μl |
| DOP amplified BAC clone |
2 μl |
| Total volume |
50 μl |
|
* SpectrumGreen™- dUTP (Vysis), SpectrumOrange™- dUTP (Vysis) or biotin-dCTP | |
Backx et al. Molecular Cytogenetics 2008 1:3 doi:10.1186/1755-8166-1-3 |
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