TABLE 1.
Oligonucleotides used as PCR primers for detection of targeted mutated ES cells
| Primer | Binding site | Orientationa | Specificityb | Sequencec |
|---|---|---|---|---|
| 15F | Exon 15 | → | 5′-TGCTGACCGAGGACTGACCACTCGACCAG-3′ | |
| 16F-wt | Exon 16 | → | wt | 5′-GAGATCTCGGAAGTGAAGATGGATGCAGAA-3′ |
| 16F-S | Exon 16 | → | SFAD | 5′-GAGATCTCGGAAGTGAATCTAGACGCGGAG-3′ |
| 16R-wt | Exon 16 | ← | wt | 5′-CCTGAATCATGTCCGAATTCTGCATCCATC-3′ |
| 16R-S | Exon 16 | ← | SFAD | 5′-CCTGAATCATGTCCGAACTCCGCGTCTAGA-3′ |
| 17F-wt | Exon 17 | → | wt | 5′-GGCGTTGTCATAGCAACCGTGATTG-3′ |
| 17F-H | Exon 17 | → | HCHWA-D | 5′-TGTTCTTTGCGCAGGACGTCGGA-3′ |
| 17R | Exon 17 | ← | 5′-ATCACCAGGGTGATGACAATCACGGTTGCT-3′ | |
| 17R-H | Exon 17 | ← | HCHWA-D | 5′-ATGGCGCCTTTGTTCGATCCGACGTCCTGC-3′ |
| 17+R | Intron 17d | ← | 5′-TGGTTCTCTCTGTGGTAGCCTTGG-3′ | |
| NeoF | Neo gene | → | 5′-ATATTGCTGAAGAGCTTGGCGGC-3′ |
a
Arrows indicate the 5′-to-3′ orientation of primers binding to genomic DNA with respect to the transcriptional direction of the β-APP gene.
b
If specificity is not indicated, primers bind unspecifically to wild-type (wt) and mutant DNA.
c
Underlined letters represent mutant-specific nucleotide exchanges.
d
The binding site of primer 17+R is located 2.0 kb upstream of exon 17.