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Table 8 The tetra-primer Amplification Refractory Mutation System-PCR primers and conditions for genotyping CARD15 SNPs.

From: Identification of single nucleotide polymorphisms in bovine CARD15 and their associations with health and production traits in Canadian Holsteins

Position

Primer Name

Sequence, 5' → 3'

Tann, °C

Amplicon size, bp

c.2886-14A>G

Forward inner primer

Reverse inner primer

Forward outer primer

Reverse outer primer

CAAGCATCCTCAAAGTTCACCATTA TG

GTGGTTGTTAGACAGCCTAGAGAGGAAT AT

CCTGGCACTAAAATGCACGTATTTTTAT

AGTTATACGTTGGAAAACTGAGGCTCAG

60°C

405bp(outer)

300 bp (allele 'A')

150 bp (allele 'G')

c.3020A>T

Forward inner primer

Reverse inner primer

Forward outer primer

Reverse outer primer

GAGGAAATTGAGAAACTCAGCCAG CA

AAACATCAGAGCAAGAGTCTGGTATG CA

CTGACAGAGGGTAAAACCTGCACAGACG

CTGTGCCAGAACAAAGGTGACCTATTG

59°C

310bp (outer)

197 bp (allele 'A')

166 bp (allele 'T')

c.4500A>C

Forward inner primer

Reverse inner primer

Forward outer primer

Reverse outer primer

AGAGACGCAAGCAGGCCCCTGGGCCG CA

CCCTGGAGACACTTGGAGAGAATGGG GG

AGCAGTGTTTAGAAATAGCCTCGCAAT

AGAGAACCCACACACATGCCCTTACTG

64°C

400bp (outer)

250 bp (allele 'A')

150 bp (allele 'C')

c.4950C>T

Forward inner primer

Reverse inner primer

Forward outer primer

Reverse outer primer

TGTATCCATTCAATATATAATACATTG GC

GGCTAGCAAATGTATTATGAGGTG AA

TTCATTTGTTTGAATTTATTTTATTGAGG

GAATGTAACACAAAGGAACATATCTACAA

59°C

341bp (outer)

226 bp (allele 'T')

169 bp (allele 'C')