Primers are designed to amplify sequences present only in the vector backbone. If a correct homologous recombination has occurred then no product should be amplified. If a product is detected it strongly suggests an incorrect targeting event or a multiple insertion event.
Not all vector primers will be present in all backbones – please refer to the tables below on which primers to use.
Please note that the absence of a product does not necessarily prove that the targeting is correct and this test should be used in conjunction with others
| Primer |
Sequence |
| VF1_F |
CTCCTTTCGCTTTCTTCCCT |
| VF1_R |
GCGAAAAACCGTCTATCAGG |
| VF2_F |
TTGATTTGGGTGATGGTTCA |
| VF2_R |
GAAATCGGCAACATCCCTTA |
| VF4_F |
GTAGCTGACATTCATCCGGG |
| VF4_R |
CTTTTCTACGGGGTCTGACG |
| VF5_F |
CGAACGACCTACACCGAACT |
| VF5_R |
AAAGATACCAGGCGTTTCCC |
| VF6_F |
CAGCATGCCTGCTATTGTCT |
| VF6_R |
TAGTTGCCAGCCATCTGTTG |
| VF7_F |
GTGGCGCTTTCTCATAGCTC |
| VF7_R |
GCCTACATACCTCGCTCTGC |
| VF8_F |
ATAATACCGCGCCACATAGC |
| VF8_R |
TTTGCCTTCCTGTTTTTGCT |
| Vector |
VF1_F |
VF1_R |
VF2_F |
VF2_R |
VF4_F |
VF4_R |
VF5_F |
VF5_R |
VF6_F |
VF6_R |
VF7_F |
VF7_R |
VF8_F |
VF8_R |
| L3L4_pZero_DTA_kan |
y |
y |
n |
n |
y |
y |
y |
y |
n |
n |
n |
n |
n |
n |
| L3L4_pZero_kan |
y |
y |
y |
y |
y |
y |
y |
y |
n |
n |
n |
n |
n |
n |
| L3L4_pD223_DTA_spec |
n |
n |
n |
n |
n |
y |
y |
y |
y |
y |
n |
n |
n |
n |
| L3L4_pD223_DTA_T_spec |
n |
n |
n |
n |
n |
y |
y |
y |
y |
y |
n |
n |
n |
n |
| R3R4_pBR_DTA+_Bsd_amp |
n |
n |
n |
n |
n |
n |
n |
n |
n |
n |
y |
y |
y |
y |
