Table 2

Protocol of DOP–PCR

Reagent	Volume (μl)	Reaction
10× high salt buffer: 200 mM Tris/HCl (pH 9.2), 600 mM KCl, and 20 mM MgCl₂.
10× low salt buffer: 100 mM Tris/HCl (pH 8.4), 100 mM KCl, and 15 mM MgCl₂.
DOP–PCR, degenerate oligonucleotide primed polymerase chain reaction.
Preamplification step
10× high salt buffer	0.5	1 minute denaturation at 94°C
2 mM dNTP	0.5	1 minute annealing at 25°C
10 μM UN1 primer	0.5	3 minute ramp from 25°C to 74°C
ThermoSequenase (4 U/μl)	0.5	2 minutes extension at 74°C
Sample	3.0	4 cycles in total
	5.0 (total)
Second amplification step add
10× low salt buffer	2.0	1 minute denaturation at 94°C
2 mM dNTP	5.0	1 minute annealing at 56°C
Water	12.2	2 minutes extension at 72°C
100 μM UN1-primer	0.3	30 cycles in total
Taq polymerase LD (5 U/μl)	0.5
	25.0 (total)