Table 1.
(A) Primers used in RT-PCR | |||||
Probe set | Primer name | Direction | AT | Primer sequence (5' – 3') | Target region |
115.m00143_at | 115.m00143F | S | 55°C | CCAAACGATACACACCCAGA | Coding |
115.m00143R | AS | 55°C | GCAATTTGAAACTTCACATCACA | Coding | |
141.m00082_at | 141.m00082F | S | 55°C | TCATTTGGAATTGTTTATGTTGG | Coding |
141.m00082R | AS | 55°C | TGCACTTCCAAAATCCGTTA | Coding | |
2.m00545_at | 2.m00545F | S | 55°C | GCTGCCATGACTAATGCTGA | Coding |
2.m00545R | AS | 55°C | AGCAACAGCAACTGGTCCTT | Coding | |
226.m00092_at | 226.m00092F | S | 55°C | GCAAACATGGGATACAGCAG | Coding |
226.m00092R | AS | 55°C | ACAAGGCGCTTGTTCAACTT | Coding | |
(B) Primers used in PCR from bisulfite treated DNA | |||||
Probe set | Primer name | Direction | AT | Primer sequence (5' – 3') | Target region |
64.m00187_s_at | EHsp5'bis* | S | 50°C | ATGAATAAGAAAGTGTGAATAATAG | Promoter |
EHsp3'bis* | AS | 50°C | AACATTAATTCCACTATTTCCTACTA | Promoter | |
EHsp1005'* | S | 50°C | TGAGTATTTAAAGGAACTTGAAG | Coding | |
EHsp3'bis* | AS | 50°C | AACATTAATTCCACTATTTCCTACTA | Coding | |
115.m00143_at | 115.mPF | S | 52°C | CTTGAAGGATAAAAAATAAGTCATATTTCTA | Promoter |
115.mPR | AS | 52°C | ATAATCTGCATGGCATATATTTCCTAATT | Promoter | |
115.m00143_at | 115.mBCF | S | 48°C | ATGTTTGTATCTTATTTTCTATTTTTAATTT | Coding |
115.mCR | AS | 48°C | GTTCCAGTACATACAATTTTACCT | Coding | |
141.m00082_at | 141.mBPF | S | 52°C | TATTGAACACTGCTCTAAATCCACT | Promoter |
141.mBPR | AS | 52°C | GTGTAGTACTGAATAAACACTTTTCTT | Promoter | |
141.mBCF | S | 53°C | TTGATATAACTAATTCACCTACATTCCAA | Coding | |
141.mBCR | AS | 53°C | GATCCTTCTCCTATTTTCTTTTCTTCT | Coding | |
97.m00140_at | 97.mPF | S | 50°C | ATTACTCCTTCCCTCTCTTCTT | Promoter |
97.mPR | AS | 50°C | CCATTRTCATTTCTCTCAACC | Promoter | |
194.m00103_at | 194.mPF | S | 50°C | GACTRCACCCAATTTTCCACCT | Promoter |
194.mPR | AS | 50°C | CCATCCCAATAAATTCCTTCTT | Promoter | |
687.m00016_at | 687.mBPF | S | 53°C | CTTTCARACACTTAACAAATCTTCTTCA | Promoter |
687.mBPR | AS | 53°C | ARATTCTCTCAAATACACTTCCCAC | Promoter | |
3.m00674_at | 3.mBPF | S | 49°C | GTTATTCAATTATACTTTATACCAAACA | Promoter |
3.mBPR | AS | 49°C | TCATTTTARTTCTTTTTTCTTATACCACT | Promoter | |
160.m00098_at | 160.mBPF | S | 50°C | CTAATAAAATTTTCTTACACCTAACTCA | Promoter |
160.mBPR | AS | 50°C | CTTCTTTATTTTTTCCATAATATTCTTTCT | Promoter | |
26.m00304_at | 26.mBPF | S | 48°C | TAAAACTARACCATTTATTCAACAATT | Promoter |
26.mBPR | AS | 48°C | RRTTCTTCTTCCATAATTTATTATTA | Promoter | |
2.m00545_at | 2.mBPF | S | 50°C | TTACTTCTCGTTATTCTATTAATATAAAC | Promoter |
2.mBPR | AS | 50°C | GATTATTTTTTGCCATCCATTTATCAA | Promoter | |
2.mBCF | S | 50°C | CTAAATTTACAAAAAAACTAATATACTCTC | Coding | |
2.mBCR | AS | 50°C | TAGAGCCACCATTACATCCATTA | Coding | |
226.m00092_at | 226.mBPF | S | 49°C | CTATTGATATTGAAATGCAAAAAACAAT | Promoter |
226.mBPR | AS | 49°C | TGATATATCAACTTAAACAAACTTACTA | Promoter | |
226.mBCF | S | 50°C | GTTCTTCTAGATTATTAAAAACCATTC | Coding | |
226.mBCR | AS | 50°C | AACACTTCCTTTATCTATTTTATTTCC | Coding |
Primers used, annealing temperatures, sequences, and their targets (gene promoters or coding regions) are listed for the (A) RT-PCR and (B) bisulfite sequencing studies. The probe set targeted, the primer name, direction of the primer, annealing temperature, primer sequence, and region of each gene targeted (coding or promoter) are shown. S refers to the sense primer; AS to the antisense primer; R represents either A or G. * indicates primers taken from Bernes, et al., 2005 [16].