DNA template	(100 ng)
5x phusion HF buffer	4 μl
10 mM dNTPs	0.4 μl
Pre-index forward primer (10 μM) ) (Table 1)	1 μl
Pre-index reverse primer (10 μM) (Table 1)	1 μl
Phusion DNA polymerase (2 U/μl)	0.2 μl
Distilled water	Up to 20 μl
Total volume	20 μl