Selection of reference genes for normalisation of real-time RT-PCR in brain-stem death injury in Ovis aries

BMC Molecular Biology

Table 1 Primer information for candidate normalisation genes.

Gene	Forward primer (5' → 3')	Reverse primer (5' → 3')	Tm (°C)^a	Amplicon size (bp)	PCR Efficiency^b
ACTB	CCAAGGCCAACCGTGAGA	AGCCTGGATGGCCACGT	59	80	1.87
GAPDH	ATGCCTCCTGCACCACCA	AGTCCCTCCACGATGCCAA	60	76	1.88
HPRT	GCTGAGGATTTGGAGAAGGTGT	GGCCACCCATCTCCTTCAT	58	94	1.95
PGK1	ACTCCTTGCAGCCAGTTGCT	AGCACAAGCCTTCTCCACTTCT	59	101	2.03
PPIA	TCATTTGCACTGCCAAGACTG	TCATGCCCTCTTTCACTTTGC	59	72	1.78
RPLP0	CCAGGCTTTAGGCATCACCA	GGCGCCTACTTTGTCTCCTGT	60	94	1.96

^aTheoretical melting temperature calculated with Primer Express software (Applied Biosystems).
^bPCR efficiencies were determined using the formula 10^-1/slope

ISSN: 1471-2199