Assisted Reproductive Technology affects developmental kinetics, H19 Imprinting Control Region methylation and H19gene expression in individual mouse embryos

Table 1 PCR primers for DNA methylation and expression analysis

Gene	Primer number	Position	Primer sequence (5' to 3')	AT (°C)	Cycle number	PCR (bp)	Sequence reference
DNA methylation of H19
ICR^{CTCF 1–2}	Primers 1						AF049091
	Sens		GATTAGATAGTATTGAGTTTGTTTGGAGT
	AntiSens		ATCAAAAACTAACATAAACCCCT	47	35
	Primers 2-Nested
	Sens	1377/	GAGAAAATAGTTATTGTTTATAGTTTT
	AntiSens	1971	CCTCATTAATCCCATAACTAT	47	35	574
ICR^{CTCF 3–4}	Primers 1						AF049091
	Sens		GGTTTTTTTGGTTATTGAATTTTAAAATTAG
	AntiSens		AAAAACCATTCCCTAAATACACAAATACC	47	35
	Primers 2-Nested
	Sens	2825/	TTAGTGTGGTTTATTATAGGAAGGTATAGAAGT
	AntiSens	3433	TAAACCTAAAATACTCAAAACTTTATCACAAC	47	35	660
Proximal promoter	Primers 1						AF049091
	Sens		TGATTGGTTAGTTTTTGAGTTTT
	AntiSens		TAATAACTAATTTAAACACTCCTCACC	50	40
	Primers 2
	Sens	4648/	GGTGTTTTGATTTGTGG
	AntiSens	5019	TAATAACTAATTTAAACACTCCTCACC	50	40	371
Expression analysis
H19	Primers						NR_001592.1
	Sens	6995/	ACATGACATGGTCCGGTGT
	AntiSens	7145	TCCCATGGTGTTAACGAAGGC
	Universal Probe 55		GGAGAGGA	54	55	69
SDHA	Primers						NM_023281.1
	Sens	1435/	CCCTGAGCATTGCAGAATC
	AntiSens	1504	TCTTCTCCAGCATTTGCCTTA
	Universal Probe 80		CCTGGAGA	54	55	70

ISSN: 1471-213X