Extended Data Table 1.
Mouse genotyping
| Mouse line | Primer 1 sequence 5' --> 3' |
Primer 2 sequence 5' --> 3' |
Denaturation | Annelaling | Elongation | Cycles | Final Enlongation | Expected bands | |
|---|---|---|---|---|---|---|---|---|---|
| Csf1riCre | Mutant allele | TCTCTGCCCAGAGTCATCCT | CTCTGACAGATGCCAGGACA | 94°C – 45sec | 60°C – 45sec | 72°C – 45sec | 30 | 72°C – 5 min | Mutant: 400 bp |
| CsflrMeriCreMer | WT allele Mutant allele |
TCGAAGCTTGCATGCCTGCA TCATTCCAGAACCAGAGC |
TCTCTGCCCAGAGTCATCCT GATCGTGTTGGGGAAGCC |
94°C – 45sec | 60°C -90sec | 72°C – 90sec | 30 | 72°C – 5 min | WT: 1Kb Mutant: 2 Kb |
| Cff1rflox | Mutant allele | GCCACCAT GTGTCCGTG CTT | ACCCAGAGCCCCCACAGATA | 94°C – 30sec | 60°C – 60sec | 72°C – 60sec | 35 | 72°C – 2 min | WT: 450 bp Floxed: 500 bp |
| Cfflr+/− | Mutant allele | GGT GGAT GT G GAAT GTGTGCG | CGTTTCTTGTGGTCAGGGTGC | 95°C – 20sec | 62°C – 20sec | 72°C – 40sec | 35 | 72°C – 5 min | Mutant: 314 |
| Flt3Cre | Mutant allele | ACGGAGTCCAGGCAACTTCC | GAAGCATGTTTAGCTGGCCC | 94°C – 30sec | 58°C – 35sec | 68°C – 60sec | 44 | 72°C – 2 min | Mutant: 400 bp |
| Myb+/− | WT allele Mutant allele | CCATGCGTCGCAAGGTGGAAC CCATGCGTCGCAAGGTGGAAC |
GTGCTTCGGCGATGTGGTAA TGGCCGCTTTTCTGGATTCATC |
94°C – 30sec | 60°C – 30sec | 72°C – 60sec | 35 | 72°C – 7 min | WT: 200 bp Mutant: 300 bp |
| Rosa.26CreERT2 | Mutant allele | GCCTGCATTACCGGTCGATGCAACGA | AATGGTGTTGCCGCGCCATCTGCCAC | 95°C – 20sec | 67°C – 20sec | 72°C – 50sec | 40 | 72°C – 2 min | Mutant: 700bp |
| Rosa26LSL-yFP | WT allele Mutant allele |
AAGTCGCTCTGAGTTGTTAT AAGTCGCTCTGAGTTGTTAT |
GGAGCGGGAGAAATGGATATG GCGAAGAGTTTGTCCTCAACC |
94°C – 40sec | 60°C – 60sec | 72°C – 60sec | 34 | 72°C – 5 min | WT: 525 bp Mutant: 300 bp |
| Rosa26LSL-tdTornato | WT alelle | AAGGGAGCTGCAGTGGAGTA | CCGAAAATCTGTGGGAAGTC | Step 1: 94°C – 20sec | 65°C – 15sec
(0.5C decrease per cycle) 60°C – 15sec |
68°C – 10sec | 10 | 72°C – 2 min | WT: 297 bp |
| Mutant allele | GGCATTAAAGCAGCGTATCC | CTGTTCCTGTACGGCATGG | Step 2: 94°C – 15sec | 60°C – 15sec | 72°C – 10sec | 28 | Mutant: 196 bp | ||
| Tnfrff11aflox | WT allele | AGTGTGCCTGGCATGTGCAGACCTT | CTGGTGGTTGTTCTCCTGGTGTCAT | 94°C – 30sec | 60°C – 30sec | 72°C – 40sec | 35 | 72°C – 5 min | WT: 256 bp |
| Mutant allele | AGTGTGCCTGGCATGTGCAGACCTT | GGCAGAACTCGGATGCACAGATTGG | Floxed: 390 bp Delta: 566 bp |
||||||
| Tnfrsf11aKoba/Cre | WT allele Mutant allele |
TGAAGGGTGACATCATCGTGGT ACTTCTCCATGGTAGCCTCC |
AATAGGGGTGGGGTGATA AATAGGGGTGGGGTGATA |
94°C – 30sec | 60°C – 30sec | 72°C – 30sec | 35 | 72°C – 5 min | WT: 530 bp Mutant: 274 bp |
| TnfrsfllaWafk/Cre | Mutant allele | AACCTGAGGATGTGAGGGACTA | GTCAAAGTCAGTGCGTTCAAAG | 95°C – 30sec | 58°C – 30sec | 68°C – 60sec | 35 | 72°C – 2 min | Mutant: 210 bp |
| VavDD/Cre | Mutant allele | GCCTGCATTACCGGTCGATGCAACGA | AATGGTGTTGCCGCGCCATCTGCCAC | 95°C – 20sec | 67°C – 20sec | 72°C – 50sec | 40 | 72°C – 2 min | Mutant: 700bp |
| CtfkCre | TTATTCCTTCCGCCAGGATG | TTGCTGTTATACTGCTTCTG | 95°C – 30sec | 55°C – 30sec | 72°C – 60sec | 35 | 72°C – 5 min | WT: 140 bp | |
| TAGTTTTTACTGCCAGACCG | Mutant: 300 bp |