Table 2.
Primers used in this study
| Primer use and target gene | Primer name | Primer sequencea (5′ → 3′) |
|---|---|---|
| Target gene amplification | ||
| fadB | DH1_fadB_SLIC_F1 | GCGAAGCAGTTGCAGCCTTTAGTAAATCATGACTCATAAGAGCTCGGTACGACCAGATCACCTTGCGG |
| DH1_fadB_SLIC_R1 | TGGACGGTCATGACGATGCTCCTGTTCGTGAGTGGGGGCGTTCGAACGGCCCATCGGGGT | |
| DH1_fadB_F1 | CTGCCATATGCTTTACAAAGGCGACACCCTGT | |
| DH1_fadB_R1 | TACAGAATTCGAACGGCCCATCGGGGTG | |
| fadM | DH1_fadM_F1 | CGCTGAATTCACAACGTAAGGTTATTGCGCTATGC |
| DH1_fadM_R1 | ATGTTCTAGACTTGAGCATCCGGCACCACAAAAC | |
| hcaT | DH1_hcaT_F1 | TACTGAATTCCCTGACGGGAGGGACTCATGGT |
| DH1_hcaT_R1 | GCTATCTAGAGGAGCAGATCCGCAAAATGCTCG | |
| l-mbp | l-mbp_SLIC_F1 | TGTGGAATTGTGAGCGGATAACAATTGCACCAACAAGGACCATAGCATATGAAAATCGAAGAAGGTAAACTGGT |
| l-mbp_SLIC_R1 | AAGGCGCTTGCCAGGCTCGTCGTTGCCATCCCGAGGTTGTTGTTATTGTTATTGTTG | |
| paaI | DH1_paaI_F1 | AGTGGAATTCGGGCGCTTCTGGAGAGCGGTTA |
| DH1_paaI_R1 | TTATTCTAGAGGCTTCACGCATCAGGCTTCTCC | |
| ptrc | Ptrc_SLIC_F1 | GTTTTGTGGTGCCGGATGCTCAAGTCTAGATATCATCGACTGCACGGTGC |
| Ptrc_SLIC_R1 | TTCCATGTTTCCTCCTGCGCAGGGAATTCCATGGTCTGTTTCCTGTGTGA | |
| ShMKS1 | MKS1_SLIC_F1(MKS2) | CGTCCAGCATCATCTGTAATCTAGACCTGCGCAGGAGGAAACATGGAA |
| MKS1_SLIC_F2 (fadM) | TTTTGTGGTGCCGGATGCTCAAGTCTAGACCTGCGCAGGAGGAAACATGGAA | |
| MKS1_SLIC_F3 (ptrc) | TCACACAGGAAACAGACCATGGAATTCCCTGCGCAGGAGGAAACATGGAA | |
| MKS1_SLIC_R1 | GCCAAGCTTGCATGCCTGCAGGTCGACTCATTTGTATTTATTAGCGATGG | |
| ShMKS2 | MKS2_SLIC_F1 | TCACACAGGAAACAGACCATGGGATCCCCTGCGCAGGAGGAAACATGTCAC |
| MKS2_SLIC_R1 | TTCCATGTTTCCTCCTGCGCAGGTCTAGATTACAGATGATGCTGGACG | |
| Mlut_09310 | Mlut_09310_C123S_F1 | TCTCCGCCGCGAGCGCCGGCTAC |
| Mlut_09310_C123S_R1 | GTAGCCGGCGCTCGCGGCGGAGA | |
| Mlut_09310_H275A_F1 | CCGCGTTCATCCCGGCCCAGGCCAACATGC | |
| Mlut_09310_H275A_R1 | GCATGTTGGCCTGGGCCGGGATGAACGCGG | |
| Mlut_09310_N306A_F1 | GCGGACGCCGGCGCCACGTCGGCCGC | |
| Mlut_09310_N306A_R1 | GCGGCCGACGTGGCGCCGGCGTCCGC | |
| Mlut_11700 | Mlut_11700_SLIC F1 | GTCATTGTCGATGCAATTCGCACCCCGATGGGCCGTTCGAACGCCCCCACTCACGAACAGG |
| Mlut_11700_SLIC R1 | TGCCTCTAGCACGCGTCTCACTATAGGGCGAATTGGAGCTCCACCGCGAGGTGACGGGG | |
| Mlut_11700_F2 | GATTCATATGACCGTCCACGAGAAGCTCGC | |
| Mlut_11700_R2 | GATTGAATTCACCGCGAGGTGACGGGGG | |
| UcfatB1 | UcfatB1_SLIC_F1 | CAACAATAACAATAACAACAACCTCGGGATGGCAACGACGAGCCTGGCAAGCGCCTT |
| UcfatB1_SLIC_R1 | ATCCGCAAGGTGATCTGGTCGTACGAGCTCTCACACACGCGGTTCAGCCGGAAT | |
| Real-time PCR | ||
| fadM | fadM_qPCR_ F1 | CCGCTACCTTGAATTTCTCG |
| fadM_qPCR_ R1 | ACGACGAAGGCGATGTTATG | |
| hcaT | hcaT_qPCR_ F1 | GCTGATGCTGGTGATGATTG |
| hcaT_qPCR_R1 | AGTCGCACTTTGCCGTAATC |
a
Underlined sequences indicate restriction sites or homology regions used for cloning purposes. Bold sequences indicate nucleotide changes from wild-type gene to generate site-directed mutations.