TABLE 5.

Design of Illumina-tagged, phase-offset ITS1F and ITS2 primers for fungal amplification

Primer direction and name	Illumina tag	Phase offset	Linker	Gene primer	Full primer sequence (5′→3′)
Forward
ITS1F-A	ACACTGACGACATGGTTCTACA		GG	CTTGGTCATTTAGAGGAAGTAA	ACACTGACGACATGGTTCTACAGGCTTGGTCATTTAGAGGAAGTAA
ITS1F-B	ACACTGACGACATGGTTCTACA	C	GG	CTTGGTCATTTAGAGGAAGTAA	ACACTGACGACATGGTTCTACACGGCTTGGTCATTTAGAGGAAGTAA
ITS1F-C	ACACTGACGACATGGTTCTACA	AC	GG	CTTGGTCATTTAGAGGAAGTAA	ACACTGACGACATGGTTCTACAACGGCTTGGTCATTTAGAGGAAGTAA
ITS1F-D	ACACTGACGACATGGTTCTACA	TAC	GG	CTTGGTCATTTAGAGGAAGTAA	ACACTGACGACATGGTTCTACATACGGCTTGGTCATTTAGAGGAAGTAA
Reverse
ITS2-A	TACGGTAGCAGAGACTTGGTCT		CG	GCTGCGTTCTTCATCGATGC	TACGGTAGCAGAGACTTGGTCTCGGCTGCGTTCTTCATCGATGC
ITS2-B	TACGGTAGCAGAGACTTGGTCT	G	CG	GCTGCGTTCTTCATCGATGC	TACGGTAGCAGAGACTTGGTCTGCGGCTGCGTTCTTCATCGATGC
ITS2-C	TACGGTAGCAGAGACTTGGTCT	TG	CG	GCTGCGTTCTTCATCGATGC	TACGGTAGCAGAGACTTGGTCTTGCGGCTGCGTTCTTCATCGATGC
ITS2-D	TACGGTAGCAGAGACTTGGTCT	ATG	CG	GCTGCGTTCTTCATCGATGC	TACGGTAGCAGAGACTTGGTCTATGCGGCTGCGTTCTTCATCGATGC