Table 4.

Some suggested human reference primers

Gene	Primer sequence	Exon location ID	Accession #T Cat. #
HMBS	GCAACTGTACCTGACTGGA TCAGGGCCATCTTCATGC	E12–13F, 60.1 E14R, 60.7	NM_000190.3 Hs. PT.58.242481164
HPRT1	CATCAAAGCACTGAATAGAAATAGTGA CCAATTACTTTTATGTCCCCTGTT	E3F, 60.5 E4R, 61.1	NM_000194 Hs. PT.53a.20881146
HPRT1	AGGTATGCAAAATAAATCAAGGTCAT TTCTCCTGAGCAGTCAGC	E1F E2R	NM_000194 Hs. PT.53a.2145446
YWHAZ	TCTGATCCCCAATGCTTCAC TGGTATGCTTGTTGTGACTGA	E8F, 60.9 E9R, 60.9	NM_003406 Hs. PT.53a.20437075

A discussion of reference gene choice can be found in Refs. [3, 10]. Primer sequences are 5′ to 3′. Catalog numbers are for IDT’s Prime Time Pre-Designed qPCR Assays. The exon number may be different for different accession numbers. The T_m values were determined using IDT’s OligoAnalyzer tool using the following final concentrations in the qPCR reaction: [primers] = 200 nM, [Na⁺] = 55 mM, [Mg²⁺] = 1.5 mM, and [dNTP] = 0.2 mM. The actual annealing temperatures were determined by performing a gradient RT-qPCR for each primer set using the methods described in the text. Primers for mouse or rat can be obtained by entering the name of the gene and desired species on the IDT website