. 2019 Feb 6;14(2):e0210952. doi: 10.1371/journal.pone.0210952

Table 2. Genus- and species-specific assays for hierarchical detection of Melampsora spp., Melampsora medusae and Melampsora larici-populina.

Assay	Targeted taxon	Primer or TaqMan probe^a	Sequence (5’ → 3’)	Tm (NN)^b	Amplicon length	GenBank accession no.
MEL40	Melampsora spp.	Melampsora40Fm	`CCTGGTACTCCAACTATCATCTTA`	60.3 °C	131 bp	MH171821-MH171846
		Melampsora40Rm	`GAAWGTGCACGCGATTGACG`	59.3 °C
		Melampsora40T (40–66)	`CCTCTGAGTGATGGCGAAGACCCTTTA`	68.1 °C
MEL100	Melampsora spp.	Melampsora100Fm	`CACGAAAGTCBCAAGTGGC`	60.6 °C	148 bp	MH171847-MH171875
		Melampsora100Rm	`GTRCAGTCATGAGGTACGATA`	59.3 °C
		Melampsora100T (66–85)	`CCTCCTCCTCCGCTCTACGC`	68.8 °C
MEL176	Melampsora spp.	Melampsora176Fm	`GCCCTTGCCGTTGCTAT`	60.7 °C	112 bp	MH171876-MH171903
		Melampsora176Rm	`GRCTCGTGCTGATCAGTC`	60.2 °C
		Melampsora176Td (37–62)	`ACCACTCACCATCCGTAYATCAACGC`	68.2 °C
MM53	Melampsora medusae	Mm53F	`ACAACCAGGTGACGGAAATC`	59.1 °C	129 bp	MH171799-MH171810
		Mm53R	`GAATCGTCCGAGGAGTCATT`	58.1 °C
		Mm53Trev (52–28)	`CTTAGGTCGTTCACCCTCTGATTCG`	64.6 °C
MM74	Melampsora medusae	Mm74Fm	`CACCATGCAAATCACCAATCAC`	58.4 °C	118 bp	MH171811-MH171820
		Mm74R	`TTTGGCTCAGCCTCAGTTTT`	58.5 °C
		Mm74T (29–57)	`TGGACCTTCTTTGCAAATCAACACAGTAT`	63.4 °C
MLP104	Melampsora larici-populina	Mlp104Fm	`CGGCCAGAAATTGTGATGGAT`	59.4 °C	104 bp	MH171781-MH171789
		Mlp104R	`TGCATAGCCTTTGTGGACAG`	59.9 °C
		Mlp104T(28–56)	`CGTATGAACCATTGATTGAAGGCTTGGAC`	65.1 °C
MLP133	Melampsora larici-populina	Mlp133Fm	`ATGGACCGGGAATATGAAC`	56.7 °C	126 bp	MH171790-MH171798
		Mlp133R	`TCGTTGATCGTATCGTGGAA`	56.0 °C
		Mlp133T(33–55)	`CGCATCAGGTGGATTTGGTGAAG`	62.9 °C

^a In parentheses: probe position within amplicon.

^b Values estimated by Oligo Analyzer V1.5.