Table 1. Oligonucleotides used for bisulfite specific PCR and pyrosequencing.
CpG site (*) Included | Details | Sequences | Annealing T (°C) |
---|---|---|---|
-244, -238 | Forward | 5′-TAGGTTTTGAGGGGTATGGG -3′ | 62 |
Reverse | 5′-[Biotin]TCAAAAATACCCCTCACACTCC -3′ | ||
Sequencer | 5′-GTTAGTGGTTTAGAAGATTT-3′ | ||
-169, -163, -161 |
Forward | 5′-[Biotin]GAGTGTGAGGGGTATTTTTGATG -3′ | 63 |
Reverse | 5′-GCAACCATAATAAACCCTACACCTTC-3′ | ||
Sequencer | 5′-AAACCCTACACCTTCTATCT -3′ | ||
-146, -119 | Forward | 5′-GAGGGGTATTTTTGATGTTTGTGT -3′ | 61 |
Reverse | 5′-[Biotin] CAACCAACCAAAAACTTCCTTAAT-3′ | ||
Sequencer | 5′-TTTAGAGATGGAGAAGAAA-3′ | ||
-72, -49, -38 | Forward | 5′-GAGGGGTATTTTTGATGTTTGTGT-3′ | 63 |
Reverse | 5′-[Biotin]CCAACAACTACCTTTATATATCCC -3′ | ||
Sequencer | 5′-TTATGGGTTTTTTTATTAAG-3′ |
CpG sites indicate nucleotide position in relation to transcription start (TSS).
“−” or “+”, indicates upstream or downstream of TSS, respectively.