Table 1 Oligonucleotide primers used to genotype the Capn2 locus
From: m-Calpain is required for preimplantation embryonic development in mice
Allele | Primer | Location | Oligonucleotide Sequence |
|---|---|---|---|
Single-Step | |||
Both | Sense | Intron 4 | 5'-GGGCCCCCATTGCCTCTTAGC-3' |
Wild-type | Antisense | Exon 7 | 5'- GGATTCCTGATGCGGATCAATTTCTGC-3' |
Mutant | Antisense | PGK-Neo | 5'-CCTCGAAGTCGAGGTCGATCC-3' |
Nested PCR | |||
Wild-Type (Diagnostic) | Sense #1 | Intron 6 | 5'-CAACATCATAAGCAACGGAGAACGC-3' |
| Â | Sense #2 | Intron 6 | 5'-GCCTGTGACAGAAGTACCACCAG-3' |
| Â | Antisense #1 | Intron 7 | 5'-CTCCTCGGCCCTCCCTGTAG-3' |
| Â | Antisense #2 | Exon 7 | 5'-GGATTCCTGATGCGGATCAATTTCTGC-3' |
Mutant (Diagnostic) | Sense #1 | Intron 6 | 5'-CAACATCATAAGCAACGGAGAACGC-3' |
| Â | Sense #2 | Intron 6 | 5'-GCCTGTGACAGAAGTACCACCAG-3' |
| Â | Antisense #1 | PGK-Neo | 5'-CCTACCCGGTAGAATTGACCTGC-3' |
| Â | Antisense #2 | PGK-Neo | 5'-GACCTGCAGGGGCCCTC-3' |
Both (Internal Control) | Sense #1 | Intron 4 | 5-GGGATGAAGGCTCCCTGTTGC-3' |
| Â | Sense #2 | Exon 5 | 5'-GGGGGTGCCACCACGGAA -3' |
| Â | Antisense #1 | Intron 5 | 5'-CCCCTAGAGGACTATTGCCTGG-3' |
| Â | Antisense #2 | Intron 5 | 5'-GATGCACCCATGCCTTAAGGAGC-3' |