Table 1 Primers and oligonucleotides used in this study
Amplicon Primers (1st PCR reaction) | |
Amplicon-TBC1D4-FWD-1 | ACACTCTTTCCCTACACGACGCTCTTCCGATCTNNCAAGGAAAATAAAGGGTCAAGTCAA |
Amplicon-TBC1D4-REV-1 | CTGGAGTTCAGACGTGTGCTCTTCCGATCCTTCCTCTTACTGGCTCTGCAG |
Amplicon Primers (2nd PCR reaction) | |
Amplicon-3A-REV-2 | CAAGCAGAAGACGGCATACGAGATCAACAAGTGACTGGAGTTCAGACGT |
Amplicon-3A-ssODN-REV-2 | CAAGCAGAAGACGGCATACGAGATGTTGTTGTGACTGGAGTTCAGACGT |
T7 Endonuclease Primers | |
T7-TBC1D4-FWD | GGCCACCATACCATCTTCACA |
T7-TBC1D4-REV | ATTTGGCTCTGCTTGTAGCC |
RT-PCR 1 kb TBC1D4 amplicons | |
TBC1D4_1kb-1-FWD | GCTTTGTGCCAACTAGCATGT |
TBC1D4_1kb-1-REV | ACTGGGTAACAGTGCGGAGG |
TBC1D4_1kb-2-FWD | AGGGCCAATAGCCAACTGAA |
TBC1D4_1kb-2-REV | TGGGTAGACTCAGCCACAATG |
TBC1D4_1kb-3-FWD | TGGTGCATGTCAGAAAGAGGT |
TBC1D4_1kb-3-REV | TCAGTTGGCTATTGGCCCTC |
TBC1D4_1kb-4-FWD | TGAAGCCAAGCAGAAGACACA |
TBC1D4_1kb-4-REV | TCCTCGTCGACTTTTGGGAA |
TBC1D4_1kb-5-FWD | TTCCAGGTTGGGCGATTTGA |
TBC1D4_1kb-5-REV | TCCCTTCTCCATCACTGCAC |
4 kb fragment flanking target site | |
TBC1D4-4kb-FWD | CCTGGAAAATCTTAATGGTGCTTC |
TBC1D4-4kb-REV | GCCTAATAAACAACAGCCTCCTG |
TBC1D4 gRNA cloning oligonucleotides and ssODN template (editing) | |
TBC1D4 gRNA-edit-FWD | CACC ATTGGTATGGGACTTTCCTA* |
TBC1D4 gRNA-edit-REV | AAAC TAGGAAAGTCCCATACCAAT |
TBC1D4 ssODN Template | CTTGAGCTGGCAATGTGAGTCCTGCATCACTAAAAGGAGAGTTCTATACACAGAAACAAATCCGTCTTCACATCAAAGCTGTCTATATTGGTATGGGCATTTTTCTAGGGCCACAA AAATGAAGGGGGATGTTAGCTTCCTTGTGAAATGATTA CTCATGTCATTTAGAACTTGCAAGAGTGCCAGGTTTTAAAATGTTT |
