Reagent	Stock concetration	Final concentration	Volume (µl)
H2O			to 100 µl
HF Buffer	5×	1×	6
dNTPs	10 mM	200 µM	0.6
Nest Primer	100 µM	30 nM	0.09
16 S-R AP28	100 µM	30 nM	0.09
Block F	100 µM	3.2 µM	0.96
Block R	500 µM	3.2 µM	0.96
Polymerase	2000 U/µl	20 U/µl	0.3
template			2–5
Total			30