ENU mutagenesis reveals that Notchless homolog 1 (Drosophila) affects Cdkn1a and several members of the Wnt pathway during murine pre-implantation development

Table 1 Primers used for this study

	Forward sequence	Reverse sequence	Tm	Size
*Nle1* sequencing primers
Nle1E1/2	CTTGACTCCTCCGAACACGAG	AAACACAGCCTGTCTGTAGGTGAG	62	500
Nle1E3	GATTAAATTTGTCGCATGGTGGTA	GTCTGTTACTTGCAACGTGAGTCC	62	475
Nle1E4	TATTTCTCCTCAGGGAATGGAGAG	CCACACTCAGTCCAGTATCTGCTT	62	377
Nle1E5/6	CTGTGTTCTCCCTCACCTCTCC	ATAGTAGGCCAAGCCGTTGCT	62	557
Nle1E7	ACAGCCTTGCTCTGCTGTTAGAA	GGACCAGCTGGACTCTTGGTATAA	62	440
Nle1E8/9	TTCCTGATTCTTGCCTTATGTCAC	AACCCTAACTAAGACAACCAAGAACAA	62	544
Nle1E10	TGGAGTTGCATGTAAGCTTGTGT	GTCACTAGCCCTAAAGATGCCATT	62	488
Nle1E11/12	CCGGCCCAGGTACCTAGCTT	ACCTACAGGTTCTCCCAGAGTCTCC	62	498
Nle1E13	ACTTGATACTTGGCAGTAGGCACA	CTCCTGCTATCCAGTGCAAGG	62	570
*Nle1* genotyping primers
Nle1 GSP	GCTGTAATGTCCTGACTGT		60	637
cDNA 1	CTGTGTCGTACTCTTCAAGGTCAT	CTGTGGAGTCATCTTCTCCATATC
cDNA 2	TCAGACGACTTCACCTTATTCCTG	CAGTCAACAGCATATACCTCATCG	62	351
Nle1 DNA	TCTCCTTCAGCTCCTTCACTGT	TCCAATGGTGGAGTATAGGGTATAA	60	341
cDNA amplification primers
1^st PCR	ATATGGATCCGGCGCGCCGTCGACT₂₄	ATATCTCGAGGGCGCGCCGGATCCT₂₄	67
2^nd PCR	(NH2)ATATGGATCCGGCGCGCCGTCGACT₂₄	(NH2)ATATCTCGAGGGCGCGCCGGATCCT₂₄	67
Eed	GTGTGACATTTGGTACATGAGGTT	ACATTTATGATGGGTCAGTGTTGT	60	148
TaqMan gene expression assay
	Assay ID	Amplicon Size (bp)
Trp53	Mm01731290_g1	119
Cdkn1a	Mm04205640_g1	80
Gapdh	Mm99999915_g1	107

ISSN: 2730-6844