Reagent	Volume (μl) per reaction	Final concentration
Water	140 μl
10 x LA buffer	20 μl
2.5 mM dNTP	20 μl	0.25 mM
10 μM primer P5	4 μl	0.2 μM
10 μM primer PE_miRCat_PCR	4 μl	0.2 μM
Template cDNA from Step 71	10 μl
TaKaRa LA Taq	2 μl	0.05 U μl−1
Mix total volume	200 μl