Table 1 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 |