. 2016 Nov 21;82(24):7102–7112. doi: 10.1128/AEM.02102-16

TABLE 3.

Primers used with the 18-plex detection system^a

cps locus type	No. of beads coupled with anti-TAG sequences	Forward and reverse primer sequences (5′–3′)	Working concn (μM)	Sensitivity (pg)	PCR product size (bp)
NCL1	54	TTAATACAATTCTCTCTTTCTCTA–C₁₂–ATTCACATAGTAACATTGCGGA	0.2	10	254
		*CAACATTGCGCAGGAAATAATA
NCL2	55	ACATCAAATTCTTTCAATATCTTC–C₁₂–TTCTTGATTATGCTGTTCTCGT	0.2	0.5	333
		*AAACACAACATCCTGTACTTCA
NCL3	56	CTTAAACTCTACTTACTTCTAATT–C₁₂–ATTCAGGAGGTATTCAACCAAG	0.2	10	370
		*AATTCACTAGCATCAACAAACG
NCL4	57	ACTTACAATAACTACTAATACTCT–C₁₂–TGCTTATTATGACTGTTGCCTT	0.2	10	293
		*ATCAGTTGATAAGGTTGCTGTT
NCL5	61	AATCTCTACAATTTCTCTCTAATA–C₁₂–CAGATGAGTCAGCAAGTAATCA	0.2	1	262
		*AGGGAAGAGTAAGATTCAAGGT
NCL6	62	CTAAACATACAAATACACATTTCA–C₁₂–TGATACGGGTACTGTTGAGTAT	0.2	0.5	220
		*ATTACTACTTCTGGTTGGGTCA
NCL7	63	CTAAATCACATACTTAACAACAAA–C₁₂–GTTGATTTATTTGCGGGACTAC	0.2	1	447
		*CAGAAAAACAATAGCAGTGACC
NCL8	64	TTCAATTCAAATCAAACACATCAT–C₁₂–AAAATTTTCACTTCACCTCGAC	0.2	20	390
		*AATCTTCCAATCAATGCTACGA
NCL9	65	TACTTAAACATACAAACTTACTCA–C₁₂–GGGTAGATACGTTCTATTTGGG	0.2	0.5	309
		*GCGACGGTATATAGAGCTATTC
NCL10	66	TCTTACTAATTTCAATACTCTTAC–C₁₂–ACAAAAATAGTAGACGGGCTTT	0.2	0.5	231
		*TTGCACGCCAAGTATAAAATTC
NCL11	67	ATCTCAATTACAATAACACACAAA–C₁₂–GCCTTAATAATGGTGGGTTTTG	0.2	1	454
		*TACCTAAAACATTTTGCCCAGA
NCL12	72	CTATCATTTATCTCTTTCTCAATT–C₁₂–GTGAGAGATTTCGGTGTAGTTT	0.2	10	380
		*GCATCAGCATACATCTTTCCTA
NCL13	73	CTTTATCAAATTCTAATTCTCAAC–C₁₂–TCTTGCTAGGTCTAATCGTAGT	0.2	0.5	179
		*CTCGCTTCCAATTAATAAACCG
NCL14	74	ACACTCATTTAACACTATTTCATT–C₁₂–AAAAGAAATGGAAAGCAGTGTG	0.2	0.5	178
		*TCTTTGCTCAGCTATTGAGTTT
NCL15	75	CATAAATCTTCTCATTCTAACAAA–C₁₂–TATGCTATTGTTACGATGTGGG	0.2	0.5	169
		*CTTGGAGAGTAAAACGATAGGG
NCL16	76	TCTCATCTATCATACTAATTCTTT–C₁₂–AGGGTTATTCTTTTTGGTGGAT	0.2	0.5	200
		*TCTTGTAAGCAGAAAATCGTGA
Chz	77	AATAACAACTCACTATATCATAAC–C₁₂–ATTTTGGGCAGTGAGTGATATT	0.2	0.5	374
		*TCAATACTTTTCTTGAACCCGA
All (thrA)	33	ACTACTTATTCTCAAACTCTAATA–C₁₂–GAAAATATGAAGAGCCATGTCG	0.3	0.5	120
		*GACAACGAACATAACAGAAACTTC

xTAG sequences and the 12-carbon amine-containing group are indicated by underlining and boldface type, respectively. * indicates that the reverse primer is biotinylated at the 5′ terminus (i.e., it indicates the beginning of the reverse primer sequence).