Table 2.
The primers set for the C57BL/6 Defa genes qPCR
| Gene group | Primers sequences | Product length, bp | qPCR annealing, ° C |
qPCR efficiency |
|---|---|---|---|---|
| Defa3 | F: GAGAGATCTGGTATGCTATTG R: AGCAGAGTGTGTACATTAAATG |
98 | 62 | 1.956 |
| Defa5 | F: TCAAAAAAGCTGATATGCTATTG R: AGCTACAGCAGAATACGAAAG |
106 | 58 | 1.982 |
| Defa20 | F: GAGAGATCTGGTATGCTATTG R: AGAACAAAAGTCGTCCTGAG |
86 | 62 | 2.125 |
| Defa21/22 | F: GAGAGATCTGATCTGCCTTTG R: CAGCGCAAAAAAGGTCCTGC |
87 | 64 | 2.079 |
| Defa23 | F: GAGAGATCTGGTATGCTATTG R: AGCAGAGCGTGTATATTAAATG |
98 | 62 | 1.376 |
| Defa24 | F: GAGAGATCTGGTATGCTATTG R: AGCAGAGCATGTACAATAAATG |
98 | 62 | 1.556 |
| Defa25 | F: GTGAAGATCTGATATGCTATTG R: ACCAGAGCATGTACATTAAATG |
99 | 58 | 1.980 |
| Defa26 | F: ATTGTAGAAAAAGAGGCTGTAC R: AGCAGAGTGTGTACATTAAATG |
81 | 62 | 1.802 |
| Defa4* | F: TGAGAGGTTTGTTATGCTATTG R: AGCAGTACAAAAATCGTATTCC |
90 | 60 | 1.992 |
Reaction cycling conditions were: one enzyme activation step at 95°C for 10 min followed by 40 cycles of 95° C, 30 s, annealing for 30 s at the specified temperature and extension at 72° C for another 30 s. Fluorescence readings were taken during the extension time.
*
Not present in C57BL/6 mice.