Gene expression profiles in canine mammary carcinomas of various grades of malignancy

BMC Veterinary Research

Table 1 Primers used for real-time qPCR

Gene symbol	Forward primer	Reverse primer	Optimum annealing temp. (°C)	Optimum annealing time (sec)
eif4b	CTTTCTGGCTGAGGATGGAG	GGGCAGGTTCCCTAGAAAAG	59	10
atf6	GTCTCCAGCCTCCTCAAGTG	GGCTCTGCTAAGGAGGGACT	64	6
tmem 85	AATGGGACTGCTGCCTACAC	TCTGGGATTGGTCTCCTCAC	62	6
hprt	AGCTTGCTGGTGAAAAGGAC	TTATAGTCAAGGGCATATCC	59	6
rps19	CCTTCCTCAAAAAGTCTGGG	GTTCTCATCGTAGGGAGCAAG	61	10

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 online access) and checked using Oligo Calculator (free on-line access) and Primer-Blast (NCBI database). Primers sequences are listed in Table 1. Hprt and rps19 genes were used as non-regulated reference genes for normalization of target gene expression [6, 7].

ISSN: 1746-6148