PCR Reaction Set-up
Step 1	Step 2
PCR1 reaction:	PCR2 reaction:
23.25 µL water (Molecular Biology grade)	19.15 µL Water (Molecular Biology Grade)
3µL 10x Taq polymerase Buffer	5 µL 10x Taq Polymerase Buffer
0.5µL Taq polymerase	0.6 µL Taq Polymerase
0.625 µL 20 µM Primer Arb1D (Table 4)	0.625 µL 20 µM Primer Arb2A (Table 4)
0.625 µL 20 µM Transposon-specific Primer (PMFLGM.GB 3a or Tn5Ext ) (Table 4)	0.625 mL 20 µM Transposon-specific Primer (PMFLGM.GB 2a or Tn5Int2) (Table 4)
1 µL 10 mM dNTPs	1 µL 10 mM dNTPs
1 µL genomic DNA, 100 ng	5 µL PCR1 reaction
30 µL Final reaction volume	30 µL Final reaction volume
PCR Reaction Settings
PCR1 Thermocycler Conditions:	PCR2 Thermocycler Conditions:
95 °C – 2 min	95 °C – 2 min
Repeat 5 cycles:	Repeat 30 cycles:
95 °C – 30 s	95 °C – 30 s
30 °C – 1 min	54 °C – 30 s
72 °C – 1 min	72 °C – 1.5 min
Repeat 30 cycles:	72 °C – 10 min
95 °C – 30 s	4 °C – Hold
45 °C – 30 s
72 °C – 1 min
72 °C – 10 min
4°C – Hold