| FRAGMENT | PRIMER NAME | PRIMER
SEQUENCE (5'>3') |
ANNEALING TEMPERATURE (C) |
| ORF15A | 4323 | gatgaccttacagacaaagcag | 54 |
| 4250 | tcttccccatccctcttctt | ||
| ORF15B | 4251 | ggagagggagagaaggaact | 50 |
| 4252 |
acgttctccctccacttctt |
||
| ORF15C | 4253 | gaggaggagcatggagaaggaga | 50 |
| 4254 | ctcccctctcccctctgttt | ||
| ORF15D | 4267 | aggagaggaagaaggagacc | touchdown 70>52 |
| 4268 | tccttcctcctcttccccctcccat | ||
| ORF15E | 4324 |
ggaggaaggagaaggggaaggggaggat |
touchdown 70>52 |
| 4325 | gtactcctctccatcctgcctttcattc | ||
| ORF15F | 4271 | aggaagaagaggggaagtatc | 54 |
| 4272 |
ccctgtgtgttagtaactgac |
||
| ORF15G | 4273 | gatctgtgaatatggcaaac | 54 |
| 4274 | gactggccataatcgggtcac |
For sequencing the repetitive region:
1. PCR with Takara La Taq, 50ml reaction, 33 cycles with annealing temp of 65; primers 4323 and 4853 (tctgactggccataatcgggtcac).
2. Gel purification. Elution in 50ml.
3. Sequencing reaction: 9ul PCR product with reverse primers only: 4272, 4268 and 4256 (cctcctcttccccctcac).
By Vivian W. Rabe and Dror Sharon