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. 2019 Nov 7;6:9. doi: 10.1186/s40575-019-0076-1

Table 3.

List of primer used for analysis of the canine CMAH gene

Amplicon Primer name Primer sequences (5′-3′) Location* Annealing (°C/s) Predicted product size (bp) Genbank Accession No., [Ref]
cDNA cloning
 Fragment 1 dCMAH-cE1aF CTGTTTTGTGCAGTTTGGCCTCTT exon 1 (5′-UTR) *55/60 1146 [30]
dCMAH-cE9R TTGTCTGATGGATGGGATTCCACA exon 9 NC_006617.3
 Fragment 2 dCMAH-cE8F CCTGAGAAGGTTGCTCTAATGA exon 8 *55/60 925 NC_006617.3
cDMAH-cE15R TGATCAAGATGTAGCGTCAGTAAAT exon 15 (3′-UTR) NC_006617.3
 Fragment 3 dCMAH-cE1aF CTGTTTTGTGCAGTTTGGCCTCTT exon 1 (5′-UTR) *55/180 1852 [30]
cDMAH-cE15R TGATCAAGATGTAGCGTCAGTAAAT exon 15 (3′-UTR) NC_006617.3
SNP discovery
  Fragment 1 dCMAH-gE1F CTCCAGGCTGCCGTCCTTCTA Promoter *55/30 227 NC_006617.3
dCMAH-gE1R CAACAGCTTTCAGTTCTTGAAT intron 1 NC_006617.3
 Fragment 2 dCMAH-gE2F GCCTGGATACTTGGAGGGAGG intron 1 *55/30 393 NC_006617.3
dCMAH-gE2R TGATCAGAGAGATGTCCTAAGT intron 2 NC_006617.3
 Fragment 3 dCMAH-gE3F AGTATTATCTCCTAATGGTTT intron 2 *55/30 369 NC_006617.3
dCMAH-gE3R AAGTCCGACTCTACACAGTTT intron 3 NC_006617.3
 Fragment 4 dCMAH-gE4F TGAGTTGGTGTTGGTCTTAAG intron 3 *55/30 483 NC_006617.3
dCMAH-gE4R TCCTAAGAAATTTGCTTAATAG intron 4 NC_006617.3
 Fragment dCMAH-gE5F TTGAGCATTCTTAGAAGCGAA intron 4 *55/30 500 NC_006617.3
dCMAH-gE5R CCTCACTCTGAGTGGTATATA intron 5 NC_006617.3
 Fragment 6 dCMAH-gE6F AATTCCTTGCTTCTTGATCAACA intron 5 *55/30 334 NC_006617.3
dCMAH-gE6R ATCTTAAAGTAACTACCTCATCTAC intron 6 NC_006617.3
 Fragment 7 dCMAH-gE7F GAAGGATTTCTTTCCAGATGAGC intron 6 *55/30 254 NC_006617.3
dCMAH-gE7R CAGATGGCTGTTATCACCCTT intron 7 NC_006617.3
 Fragment 8 dCMAH-gE8F GGACATAAGTGATGCTTCTCTA intron 7 *55/30 445 NC_006617.3
dCMAH-gE8R GGCGCTAAACCACTAAGCCAC intron 8 NC_006617.3
 Fragment 9 dCMAH-gE9F TTCCTGTGTTAGTCTATCCAT intron 8 *55/30 386 NC_006617.3
dCMAH-gE9R AAGCTCCGTGCTCTCAGGCAG intron 9 NC_006617.3
 Fragment 10 dCMAH-gE10F TAAGTGGATAGGATTGTGAAG intron 9 *55/30 373 NC_006617.3
dCMAH-gE10R CCTACAAATAAGGACTTGTCA intron 10 NC_006617.3
 Fragment 11 dCMAH-gE11F GATAAGAGAACTTTCCTGTAT intron 10 *55/30 433 NC_006617.3
dCMAH-gE11R TCCGTGAAGCAGTTGGTAGCG intron 11 NC_006617.3
 Fragment 12 dCMAH-gE12F CATTGCTATCAATTAAGGCTG intron 11 *55/30 360 NC_006617.3
dCMAH-gE12R GTAAAGTAAGGTGTTAACAAT intron 12 NC_006617.3
 Fragment 13 dCMAH-gE13F AGCCTGTCATATCTACTCCAT intron 12 *55/30 393 NC_006617.3
dCMAH-gE13R GAAATGAAGCCCTAATGGTGG intron 13 NC_006617.3
 Fragment 14 dCMAH-gE14F CAGTATGGAAGCACCATCTCT intron 13 *55/30 353 NC_006617.3
dCMAH-gE14R ATGTTCTTGCAATGTTAGCTC intron 14 NC_006617.3
mRNA expression
 CMAH dCMAH-cE2F TGTGCAAGCATCAAGGAGGC exon 2 *55/30 342 NC_006617.3
dCMAH-cE4R AGGCAGGACCTATTAACCAG exon 4 NC_006617.3
 GAPDH dGAPDH-cE1/2 CACAGTCAAGGCTGAGAACG exon 1/2 *55/30 101 [37]
dGAPDH-cE2 CACCAGCATCACCCCATTT exon 2 [37]

Number of exon is detemined by comaparison between Ac. No. LC382414 and NC_006617.3 in this study

NC_006617.3: Canis lupus familiaris breed boxer chromosome 35, CanFam3.1, whole genome shotgun sequence

d dog, E: exon, F Forward primer, R reverse primer, number exon number, c designed, g designed from genomic sequence in primer name