TABLE 3.
Primer sequences used in this study
| Primer | Function | Reaction or coordinatesb (plate/well) | Sequence (5′–3′) |
|---|---|---|---|
| olj376 | Preparing Tn-seq libraries (essential genome) | PCR-1 | GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTGGGGGGGGGGGGGGGG |
| olj510_mariner (mariner-1) | PCR-1 | ACTCACTATAGGAGGGCGGGAATCATTTGAAGGTTGGTAC | |
| BC_a | PCR-2 | CAAGCAGAAGACGGCATACGAGATxxxxxxGTGACTGGAGTTCAGACGTGTG | |
| olj511_mariner (mariner-2) | PCR-2 | Biotin-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTNNNNNGTGTCAGACCGGGGACTTATCAG | |
| VT1169_0386-F | Validating ordered library | 29/H10 | GACGCTTCCTGATGAACAAAAGTGC |
| VT1169_0386-R | 29/H10 | CCACCATCATCTTCTTGCTCGTAGG | |
| VT1169_1989-F | 14/H12 | GTAAAGTCTGCGTTAGAATAGCGAGAAATCC | |
| VT1169_1989-R | 14/H12 | GTCACCCCATACAACGGCAC | |
| VT1169_0152-F | 37/B10 | GGTCCATAACCCTCAATCTCATCAGC | |
| VT1169_0562-R | 34/G5 | CGGACATAAACTGCCAAATCCAGC | |
| VT1169_1175-R | 34/E2 | ACGCAAATTCGTTAGCAACTACTGC | |
| VT1169_1449-R | 37/B4 | GTTTTGTGCGAATGGCGTTTGC | |
| VT1169_1462-R | 18/F3 | GACGCTAACGGTGAGCTTTCC | |
| VT1169_1954-F | 37/H12 | GACCCCGCCTTCAATATCATTTGG | |
| mariner-2-check | GTGTCAGACCGGGGACTTATCAG |
a
The primer is named after the Illumina barcode that is used (e.g., the x's in the sequence for BC01 are replaced with Illumina barcode 1).
b
Coordinates in the ordered library from which individual colonies were isolated.