Supplementary Table 1. Primer pairs used in PCR experiments for
determination of Demodex mitochondrial genome sequences.
Primer Pair
PCR Product Information
F:
R:
F:
R:
Amplified initial fragment of DF 12S rRNA gene
Anneal: Step-down 54  51˚C; Product: ~520bp
Long-range PCR to amplify DF mtDNA based on
initial 12S gene fragment
Anneal: 63˚C; Product: ~14kb
Amplified initial fragment of DB ND5 gene
Anneal: Step-down 52  49˚C; Product: ~500bp
Long-range PCR to amplify DB mtDNA based on
initial ND5 fragment.
Anneal: 68˚C; Product: ~14kb
DF single-mite sequencing
Anneal: 68˚C; Product: 630bp
DF single-mite sequencing
Anneal: 60˚C; Product: 687bp
DF single-mite sequencing
Anneal: 59˚C; Product: 696bp
DF single-mite sequencing
Anneal: 58˚C; Product: 789bp
DF single-mite sequencing
Anneal: 59˚C; Product: 776bp
DF single-mite sequencing
Anneal: 60˚C; Product: 598bp
DF single-mite sequencing
Anneal: 60˚C; Product: 744bp
DF single-mite sequencing
Anneal: 60˚C; Product: 660bp
DF single-mite sequencing
Anneal: 59˚C; Product: 769bp
DF single-mite sequencing
Anneal: 60˚C; Product:579bp
DF single-mite sequencing
Anneal: 57˚C; Product: 754bp
DF single-mite sequencing
Anneal: 58˚C; Product: 757bp
DF single-mite sequencing
Anneal: 60˚C; Product: 786bp
DF single-mite sequencing
Anneal: 60˚C; Product: 792bp
DF single-mite sequencing
Anneal: 60˚C; Product: 766bp
DF single-mite sequencing
Anneal: 59˚C; Product: 581bp
DF single-mite sequencing
Anneal: 59˚C; Product: 790bp
DF single-mite sequencing
Anneal: 60˚C; Product: 735bp
DF single-mite sequencing
Anneal: 60˚C; Product: 778bp
DF single-mite sequencing
AGAGTGACGGGCGATRTGTRCA
WWRRAIHRGGATTAGATACCCT
CTCTTCCTTTTCTAAAAATCTCCTCAATTCCCAAAAC
TTGAAACTTGATTTTGTTGGCGGTTTAAAGG
F: CAAATCCACAAAAAACCCCC
R: TTGGCTATGGCTGCTCCTACTC
F: AACTGTCTAAGAGTAGAAAAAAGCAATCACCC
R: AGGATTTTTAAGTTTGTTTATTTTATGTGGTTTAGG
F: ACTAACCACAAAGACATTGGAACA
R: GAAAGAGCTTCTGAAGTTTCGGTC
F: CCCTGGACACTCCGTAGATTTAAT
R: AGTGTTACGTCTAGGGAGGAGTTA
F: ATGAGCCCACCACATATTTACAGT
R: AGTTGGCATTAGGATTGAATGAAGA
F: ACATTTTATAGGACTAAACGGCCT
R: GTTTAGTCGTCCTGGGATAGAGTC
F: ACTGAACTTACTCCTACCCAGAAC
R: TGTTGGGTCTTATGTTGGATGAGT
F: CAATTTGACCCATCGAACTCCTTC
R: TGTAGGCTTGTAAGATTGCTACCA
F: AACCCCAAACTTACTTGTACCCTT
R: AGGGCAATAGTTTACAGTTCTGGT
F: TCAGTAGCCATTTACGGAAACTGA
R: GAATCCACATTCGAAGGGTGAATT
F: AGCTGTATGATATTGGCATTTCGT
R: CACGAGTTGATTGGGCCTAAATTT
F: ATGTAGCCTTCTCTTTGGGGAAAT
R: TAGAGGTATCGGAAGGTGTCTCTT
F: AGAAATTTGGATTAGGGGAATTAACT
R: CAGCAGTTTCGGTTAAGGGATTAA
F: TCTCCTCAATTCCCAAAACTTTCA
R: TGTGGATTTGGTTTTAGTGGATTTGT
F: TATAGACCAAAAGCTCCAACCACA
R: CTGGTATTATGGGGTTGGTTGAGA
F: CGTAATGCACTCCCTTAAACCAAG
R: TTTTGGCTAATCGATTTGGGGATG
F: AAGAAGAAACAGGAGTAGGAGCAG
R: TATGGCTTTGTAAGAGGGGTTTCT
F: CCAACTTCATACCTACACAACCAC
R: TTTGTTTTGTCTGCGAAGTTCTGA
F: ACCACGCATAATTAAAACTCTCCG
R: TGTTTTGGTTATGGGGAATTGAGG
F: GCATATAAACCCCAGCTCTCAAAC
R: TTTGGGAGTAAAAGGAGTGCTTCT
F: TTCTTCCCCTTCCACGAACTAAAT
R: TTTAGTGCTTTTGAGTGCAGTTCC
F: CCCTCTGTGAATTATTTCAATCGCA
R: GAAAGTGGATGGAAAAGAAACGGT
F: TCCCACACATGAATATCAGGGTTT
R: ACGGTTCTTCGATGACTTGTTTTC
F: TATTTGCTTACGCTATCCTACGCT
R: TTGTTTAGCTGAAATAAGCCGCTC
F: CAGTACACCACGAATAACCACAAA
R: TTTGGGAGAGCGTAAAATTTTGGG
F: AATAAATTAGCAATCCCCATAAAGG
R: TGAGTGGAGTAATGGGTGCTTTTA
F: AAAATCAAAAGCCAACATCGAGGT
R: AGAGCGAAATTAGTTGAGATAACTCT
F: AAACACCACTCCCAAGCTACTAAT
R: TGAGGGAAGAGTTTGGAAGATGAA
F: GGCTTGTCTTCAAACTCCTGAATC
R: TGGTGGGATTCCTGACAATGATAA
F: ACTCTGTTCAATCTACTTCCTCACC
R: TGTTCCAATGTCTTTGTGGTTAGT
F: TTATCATTGTCAGGAATCCCACCA
R: TGGAAAATTTTGCCAGGTTGTCTT
F: CTCTTCGAAAGTCTAACCTTCCCT
R: ACTTGTTCTGGTTTAATTTTCGGGT
F: TCATAATCTTCTCCCTCCACATCG
R: ATTGCGAATACTGCTCCTATGGAT
F: GCCACAATAGTAATCGCAATTCCA
R: TTGGTTCATGTTTTGTTGGAGAGG
F: CTACTACCCAGACACCAACAATGA
R: ATTCCGCATAATTCTGAGCATTGG
F: GCTTAATCACTACCAGAAACGACG
R: TTTGTTGGTGAGTGTTAGTGATGC
F: CATCCTAAACTTCAACCCAAACCC
R: TGATGATGTATGCCTGTAGGATGG
F: AACTTCATACCCCACCTTAATCCC
R: TTCTTGATGAAAGGAGGAGGATGG
F: TCCCTAAGTCCTGCAATAGACCTA
R: GAGGTTTTGGATGGGTTGAAGATG
F: CTTCAACATCTCCCCAACAAAACA
R: TCCTTATCTTGTTTCTTTCCCCGA
F: AGAGATGAAACATTTTAACTTGCA
R: CGGAAGATGCTTCTTGTAAACCTC
F: ACTTATAGATTCTAAATCCATCGTACT
R: TTTAAGCAGGGTTGGGTGATTTTG
F: GCCCCTCTGTAAAACCTTTAAACC
R: GGGTTTTACCTGTTGTGGTTCTTT
F: ACTCTTCAAACTCACAAATCCCCT
R: GATGAAGGTTTGTTTTGGTAGGCA
F: AAGGCATACCAAACAAAGAACCAC
R: GGTTGGGATGGTTTAGGGATTACT
F: TGAACCAAAGAAGAAACCGGAGTA
R: TTGTTGGTATTGGTGGATTTGTGG
F: ACCAAACCATCACTCAAAACAAAA
R: ATGTGATTTGAGGGTTATGGTTGC
F: CGCATAACCAAAACTCTACGAGTC
R: TTTGTTTTGGTGATGGGTAGTTGG
F: CCTCAATCCAAATCCCAACACAAA
Anneal: 60˚C; Product: 746bp
DF single-mite sequencing
Anneal: 60˚C; Product: 703bp
DF single-mite sequencing
Anneal: 60˚C; Product: 623bp
DF single-mite sequencing
Anneal: 59˚C; Product: 721bp
DF single-mite sequencing
Anneal: 56˚C; Product: 704bp
DF single-mite sequencing
Anneal: 58˚C; Product: 765bp
DF single-mite sequencing
Anneal: 60˚C; Product: 794bp
DF single-mite sequencing
Anneal: 60˚C; Product: 666bp
DF single-mite sequencing
Anneal: 59˚C; Product: 777bp
DF single-mite sequencing
Anneal: 60˚C; Product: 742bp
DB single-mite sequencing
Anneal: 60˚C; Product 799bp
DB single-mite sequencing
Anneal: 60˚C; Product: 733bp
DB single-mite sequencing
Anneal: 60˚C; Product: 784bp
DB single-mite sequencing
Anneal: 60˚C; Product: 680bp
DB single-mite sequencing
Anneal: 60˚C; Product: 604bp
DB single-mite sequencing
Anneal: 60˚C; Product: 712bp
DB single-mite sequencing
Anneal: 60˚C; Product: 723bp
DB single-mite sequencing
Anneal: 60˚C; Product: 791bp
DB single-mite sequencing
Anneal: 59˚C; Product: 796bp
DB single-mite sequencing
Anneal: 55˚C; Product: 754bp
DB single-mite sequencing
Anneal: 56˚C; Product: 789bp
DB single-mite sequencing
Anneal: 60˚C; Product: 787bp
DB single-mite sequencing
Anneal: 60˚C; Product: 634bp
DB single-mite sequencing
Anneal: 60˚C; Product: 709bp
DB single-mite sequencing
Anneal: 60˚C; Product: 767bp
DB single-mite sequencing
Anneal: 58˚C; Product: 652bp
DB single-mite sequencing
Anneal: 60˚C; Product: 788bp
DB single-mite sequencing
2
R: GAAGCAGAATGAAGTCTTTGGACA
F: ACCAATGAACTAACCGTCCCAATA
R: TGGATTGTTGAGTTTGAGGCTTTG
F: CCAAAGCATCAAACCTTACCACAA
R: ATTTGTCCTCATGGGAGTACGTAG
F: CATTCAACCGGAGCTTCACTATTC
R: GTTCTTCAATTACCTGGGCTCCTA
F: CCAACCCACTATCAACACCAAATC
R: TTTTAGAGGGTGAGTCTGAGTTGG
F: CAAGAACATCCCTCCAAAACACAA
R: TGGTAATAAGCGGTTTGGTCCTAA
F: AAACTTTCTCCTTCTTGCTCAACC
R: AGGCTAAAGTGTTCTTAAAGACGA
F: ACCAACATCGAGGTAACAAACTCA
R: TTGTATAAGGCAGTGAGGATTGGT
F: TGAAGCTTAACCATCAGCCCTTAA
R: TGAGAGAGTTGTTGGGTATTTGGT
F: GGAATCTCCTCCAATTCCTGACTA
R: AGAATGCTTGAGTTGAGTAGGGTT
F: ACCCTCAACATCCAATCTCTAACA
R: GGTTGGTTGATATGATTCATCGCA
F: TAGTAGAAACAAGCCCATGACCAA
R: TAGGTCTATTGCAGGACTTAGGGA
F: TCAGTAGCCATTTACGGAAACTGA
R: CGTGAAGTCCAGTTGTCATGAAAA
Anneal: 59˚C; Product: 740bp
DB single-mite sequencing
Anneal: 60˚C; Product: 730bp
DB single-mite sequencing
Anneal: 60˚C; Product: 795bp
DB single-mite sequencing
Anneal: 60˚C; Product: 784bp
DB single-mite sequencing
Anneal: 60˚C; Product: 646bp
DB single-mite sequencing
Anneal: 60˚C; Product: 749bp
DB single-mite sequencing
Anneal: 58˚C; Product: 779bp
DB single-mite sequencing
Anneal: 60˚C; Product: 798bp
DB single-mite sequencing
Anneal: 60˚C; Product: 715bp
DB single-mite sequencing
Anneal: 59˚C; Product: 764bp
DB single-mite sequencing
Anneal: 59˚C; Product: 799bp
DB sequence identity confirmation
Anneal: 60˚C; Product: 308bp
DF sequence identity confirmation
Anneal: 60˚C; Product 304bp
Forward and reverse primers that were used together successfully in PCR
experiments are listed, along with relevant information for each such pair: purpose
of the experiment, annealing temperature used for amplification, and the resulting
product size. DF = D. folliculorum; DB = D. brevis.
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