Table 1. Primers used for PCR screening of AAVS1 integration.
| PCR screen | Forward primer sequence
(5’ → 3’) |
Reverse primer sequence (5’ → 3’) | Annealing
temperature |
|---|---|---|---|
| 5’ integration screen
(Outside AAVS1 Left Arm Homology – CAG promoter) |
TCCCCTCTTCCGATGTTGAG | TGGGCTATGAACTAATGACCCCG | 64°C |
| 3’ integration screen
(IRES/Puromycin – Outside AAVS1 Right Arm Homology) |
AGCGTATTCAACAAGGGGCT | ACCCCGAAGAGTGAGTTTGCC | 62°C |
| Biallelic targeting screen (Inside
AAVS1
Left Arm Homology – Inside AAVS1 Right Arm Homology) |
ATGCCGTCTTCACTCGCTGG | GGGGCTTTTCTGTCACCAATCC | 64°C |