Volume 18, Number 2—February 2012
Dispatch
Phylogeography of Francisella tularensis subsp. holarctica, Europe
Table
Melt-MAMA primers targeting canonical SNPs for 6 new phylogenetic branches in a study of Francisella tularensis subsp. holarctica, Europe*
SNP | SCHU† S4 position | SNP state, D/A‡ | Primers, 5′ → 3′§ | Con, µM¶ | Tm, °C |
---|---|---|---|---|---|
B.33 | 78,382 | T/C | A: ATTGCTACTTCTATTTACGCCAACAG | 0.20 | 74.3 |
D: GGGGCGGGGCGGGGCATTGCTACTTCTATTTACGCCAAGAA | 0.20 | 79.0 | |||
C: TGTGAACAACCAAGTTGGCTT | 0.20 | ||||
B.34 | 766,614 | A/G | A: GTAGCGAGCATTATTTGCTGGTTC | 0.40 | 69.2 |
D: GGGGCGGGGCGGGGCTAGCGAGCATTATTTGCTGGGTT | 0.20 | 78.6 | |||
C: ATAAAACTATAAATTTACATAAAATGAAAACTTCTC | 0.20 | ||||
B.35 | 239,479 | A/C | A: GCCTTAATCTAGTATTTTCGCTTATCTCC | 0.40 | 70.3 |
D: GGGGCGGGGCGGGGCGCCTTAATCTAGTATTTTCGCTTATCACA | 0.20 | 75.5 | |||
C: CGGGCTCTAAAATAAGATTTAAGTTAGTAAGT | 0.20 | ||||
B.36 | 1,599,292 | A/C | A: TATTATAGTTTCTAAAAACAGTCTAATTAATTTTG | 0.60 | 69.0 |
D: GGGGCGGGGCGGGGCTATTATAGTTTCTAAAAACAGTCTAATTAATTGTT | 0.20 | 73.9 | |||
C: GTTCGACCATGACTACAGTGTTG | 0.20 | ||||
B.37 | 318,602 | T/C | A: AACATTTTAGGAACTCTACGATGATAAACTTAAC | 0.20 | 69.7 |
D: GGGGCGGGGCGGGGCCATTTTAGGAACTCTACGATGATAAACTTGAT | 0.20 | 75.9 | |||
C: GAAATATCTCAATGAAATCTAATTTAACTAAAATCAC | 0.20 | ||||
B.38 | 166,885 | C/T | A: ATGCCATCAGCCATTTACTACTCACA | 0.20 | 73.7 |
D: GGGGCGGGGCGGGGCCCATCAGCCATTTACTACTCCCG | 0.20 | 80.1 | |||
C: CTTCCCTGATTTTCTAAGTTCTGCTTG | 0.20 |
*Melt-MAMA, melt-mismatch amplification mutation assay; SNP, single nucleotide polymorphism; con, concentration; Tm, melting temperature for ancestral and derived Melt-MAMA amplification products.
†SCHU strain GenBank accession no. NC_006570.
‡SNP states are presented according to their orientation in the SCHU S4 reference genome (AJ749949.2): D, derived SNP state; A, ancestral SNP state.
§D, derived allele primer; A, ancestral allele primer; C, common primer; primer tails and antepenultimate mismatch bases are in lower case.
¶Final concentration of each primer in Melt-MAMA genotyping assays.
1These authors contributed equally to this article.