Selection of ovine housekeeping genes for normalisation by real-time RT-PCR; analysis of PrPgene expression and genetic susceptibility to scrapie

Table 1 Primers sequences and real-time PCR amplification parameters

Gene	Forward & reverse primers 5' → 3'	[C] ^a	Amplicon size (bp)	Tm ^b (°C)	Slope	R^2c	E ^d
ACTB	ATGCCTCCTGCACCACCA GCATTTGCGGTGGACGAT	300	125	85	-3.597	0.999	1.897
YWHAZ	TGTAGGAGCCCGTAGGTCATCT TTCTCTCTGTATTCTCGAGCCATCT	100	102	79	-3.335	0.988	1.995
RPL19	CAACTCCCGCCAGCAGAT CCGGGAATGGACAGTCACA	200	76	83	-3.342	0.992	1.992
GAPDH	ATGCCTCCTGCACCACCA AGTCCCTCCACGATGCCAA	100	76	84	-3.485	0.991	1.936
G6PDH	TGACCTATGGCAACCGATACAA CCGCAAAAGACATCCAGGAT	300	76	81	-3.363	0.965	1.983
SDHA	CATCCACTACATGACGGAGCA ATCTTGCCATCTTCAGTTCTGCTA	200	90	82	-3.643	0.992	1.881
PrP	GCCAAAAACCAACATGAAGCAT TGCTCATGGCACTTCCCAG	300	95	83	-3.338	0.995	1.993

^a Primer concentrations in nM
^b Theoretical amplicon melting temperature calculated with Primer Express software (Applied Biosystems)
^c Correlation coefficient
^d PCR efficiency

ISSN: 1746-6148