Table 1.
Primers Used for Genotyping and Determination of Cre-Mediated Recombination by PCR
Allele | Strand | Sequence | Amplicon size, bp |
---|---|---|---|
Genotyping | |||
CD19Cre | Forward | 5′-GCGGTCTGGCAGTAAAAACTATC-3′ | 100 |
Reverse | 5′-GTGAAACAGCATTGCTGTCACTT-3′ | ||
CD19WT | Forward | 5′-CCTCTCCCTGTCTCCTTCCT-3′ | 500 |
Reverse | 5′-TGGTCTGAGACATTGACAATCA-3′ | ||
Fabp1 | Forward | 5′-TGGACAGGACTGGACCTCTGCTTTCC-3′ | 200 |
Reverse | 5′-TAGAGCTTTGCCACATCACAGGTCAT-3′ | ||
HprtWT | Forward | 5′-TTTCTATAGGACTGAAAGACTTGCTC-3′ | 200 |
Reverse | 5′-CACAGTAGCTCTTCAGTCTGATAAAA-3′ | ||
HprtCre | Forward | 5′-GCGGTCTGGCAGTAAAAACTATC-3′ | 100 |
Reverse | 5′-GTGAAACAGCATTGCTGTCACTT-3′ | ||
Notch2[ΔPEST]COIN | Forward | 5′-CCGGGCCGCGACTGAAACCCTAG-3′ | 330 |
Reverse | 5′-CCACCACCTCCAGGAGTTGGGC-3′ | ||
Notch2tm1.1ECan | Forward Nch2Lox gtF | 5′-CCCTTCTCTCTGTGCGGTAG-3′ | WT = 310 Notch2tm1.1ECan = 400 |
Reverse Nch2Lox gtR | 5′-CTCAGAGCCAAAGCCTCACTG-3′ | ||
Notch2WT | Forward | 5′-GCTCAGACCATTGTGCCAACCTAT-3′ | 100 |
Reverse | 5′-CAGCAGCATTTGAGGAGGCGTAA-3′ | ||
RosaNotch1 | Forward | 5′-GGAGCGGGAGAAATGGATATG-3′ | WT = 600 RosaNotch1 NICD = 250 |
Reverse WT | 5′-AAAGTCGCTCTGAGTTGTTATTG-3′ | ||
Reverse RosaNotch1 NICD | 5′-GCGAAGAGTTTGTCCTCAACC-3′ | ||
LoxP recombination | |||
Notch2[ΔPEST]COIN | Forward | 5′-GTACTTCAGCACAGTTTTAGAGAAC-3′ | Not recombined and not detected Recombined = 250 |
Reverse | 5′-GTGAGTCACCCGCCGGATGTC-3′ | ||
RosaNotch1 | Forward | 5′-TTCGCGGTCTTTCCAGTGG-3′ | Not recombined = 500 Recombined = 300 |
Reverse absent loxP recombination | 5′-AGCCTCTGAGCCCAGAAAGC-3′ | ||
Reverse present loxP recombination | 5′-GCCGACTGAGTCCTCGCC-3′ |
NICD, Notch intracellular domain; WT, wild type.