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. 2021 Aug 5;11:15962. doi: 10.1038/s41598-021-95402-y

Table 1.

Anaplasma marginale, A. ovis, A. phagocytophilum, and GADPH gene (internal control) RPA primers and probes*.

Target Seq Primer Length Tm GC% Product size (bp) Sequence
A. marginale Am3L_msp4 33 76.63 54.55 103 ACGAAGTGGCTTCTGAAGGGGGAGTAATGGGAG
Am3R_msp4 30 72.99 50 [5′Label-Biotin] GACTCACGCATGTCGAACGAGGTAACAGAA
Am3_probe 39 70.4 56.4 [5′-Label-FAM] TAGCTTTTACGTGGGTGCGGCCT[THF]CAGCCCAGCATTTCC[3′-block-C3spacer]
A. phagocytophilum ApL_msp4 30 75.14 60 202 TGCGGCCGCAGTATGTGCCTGCTCCCTTTT
ApR_msp4 30 62.31 40 [5′Label-Biotin] GTTATAACCTTTTACGTAAGATCCTCCCCT
Ap_probe 52 68.2 44.2 [5′Label-FAM] TGATGCGTCTGATGTTAGCGGTGTTATGAACGG[THF]AGCTTTTACGTAAGTGGT [3′block-C3spacer]
A. ovis Ao2L_msp4 35 67.75 45.71 184 AGAGACCTCGTATGTTAGAGGCTATGACAAGAGTG
Ao2R_msp4 35 68.57 48.57 [5′Label-Biotin] CCTTCTGTAGCTTGCTTCTAGTTCCACTCTAGCTC
Ao2_probe 50 65.2 38 [5′Label-FAM] AAATCCGGCTACACTTTTGCTTTCTCTAAGAA[THF]TTACTCACATCTTTCGA [3′block-C3spacer]
GAPDH gene GAPDH2_L 30 72.53 50 168 CTGAGACAAGATGGTGAAGGTCGGAGTGAA
GAPDH2_R 27 72.82 51.85 [5′Label-Biotin] TTGCCGTGGGTGGAATCATACTGGAAC
GAPDH_Probe 51 70.3 51

[5′Label-DIG]

CTGGCAAAGTGGACATCGTCGCCATCAATGACCC[THF]TTCATTGACCTTCACT[3′block-C3spacer]

*Primers, probes, and the amplification kit are protected by a patent application.