Five markers useful for the distinction of canine mammary malignancy

BMC Veterinary Research

Table 2 Primers used for real-time qPCR

Gene symbol	Forward primer	Reverse primer	Optimum annealing temp. (°C)	Optimum annealing time (sec)
hprt	AGCTTGCTGGTGAAAAGGAC	TTATAGTCAAGGGCATATCC	59	6
rps19	CCTTCCTCAAAAAGTCTGGG	GTTCTCATCGTAGGGAGCAAG	61	10
serhl	TTGGTGCTAGACACGCTGAG	CTGCAGAAAGGCACTGATGA	61	5
zfp37	GCGGAATGGGAACAACTAGA	ATGTCTGGTTTGGGAGCTTG	61	8
mipep	CCCTGAGAAAGGCAGACTTG	AGCCACTCTGCACAAGGAAT	60	5
relaxin	AAGTTGTGCCATCCTCCATC	CCAGACCGTGTTGCTATCCT	60	7
magi3	AGGTGACATTGGGAAAGACG	CAGCCCCATGTTGTACTCCT	60	8

Primers sequences used in this study and their annealing optimal temperature and time. The mRNA sequences of key genes were obtained from NCBI database. Primers were designed using PRIMER3 software (free on-line access) and checked using Oligo Calculator (free on-line access) and Primer-Blast (NCBI database). hprt and rps19 genes were used as non-regulated reference genes for normalization of target gene expression [6, 7].

ISSN: 1746-6148