. Author manuscript; available in PMC: 2021 May 1.

Published in final edited form as: Fungal Genet Biol. 2020 Mar 3;138:103364. doi: 10.1016/j.fgb.2020.103364

Table 3:

Fusion of three fragments

PCR mixture			PCR program
Fragment 1	5’arm (from 1^st PCR)	A μL	95°C, 30s
Fragment 2	NAT, NEO or HYG(from 1^st PCR)	B μL	95°C, 15s	20 cycles
Fragment 3	3’arm (from 1^st PCR)	C μL	50°C, 4 minutes
Enzyme	2X Phusion Flash PCR Master Mix	12.5 μL	72°C, X s
H₂O		12.5-A-B-C μL	72°C, 5 minutes
TOTAL		25 μL	20°C, ∞