. 2020 Feb 26;25(5):1054. doi: 10.3390/molecules25051054

Table 1.

Primer sequences of target genes.

Reference Sequence (RefSeq) mRNA	Gene Symbol	Sense and Anti-Sense Sequences		Length of the Amplicon (bp)
NM_001243084	HIF1A	sense	TTGGCAGCAACGACACAG	169 bp
NM_001243084	HIF1A	anti-sense	GCAGGGTCAGCACTACTTC	169 bp
NM_004958	MTOR	sense	TGCCTTCACAGATACCCAGTA	171 bp
NM_004958	MTOR	anti-sense	AGACCTCACAGCCACAGA	171 bp
NM_000546	P53	sense	TCAACAAGATGTTTTGCCAACTG	118 bp
NM_000546	P53	anti-sense	ATGTGCTGTGACTGCTTGTAGATG	118 bp
NM_016539	SIRT6	sense	CTCCTCCGCTTCCTGGTC	119 bp
NM_016539	SIRT6	anti-sense	TTACACTTGGCACATTCTTCC	119 bp
NM_002046	GAPDH	sense	CCCTTCATTGACCTCAACTACATG	115bp
NM_002046	GAPDH	anti-sense	TGGGATTTCCATTGATGACAAGC	115bp

Efficiency and specificity of primers were tested making standard curves with fivefold serial dilutions of a cDNA obtained from a pool of all samples. The first dilution was the two-fold diluted cDNA. All reactions were run in duplicate. All samples were analysed in duplicate and averaged. The relative expression of the target gene was normalized to the level of GAPDH in the same cDNA. Samples were analyzed by the 2^−ΔΔCt method, as described by Livak and Schmittgen [40].