Target bisulfite sequencing of human: Post-mortem Esophagus - Mucosa
A two-step PCR approach on bisulfite-converted DNA was adopted to generate target bisulfite sequencing libraries. A singleplex PCR was performed for each target using bisulfite-specific primers for APC2 (Fw:GGTTYGGGTTTTTGGTGA; Rev:CCAACCCAACCTCATTACTA), DNER (Fw:TAGTAGTAATTAGATATTAAGGAATGGAAT; Rev:TCACCTAAATCATAAAAACTCCTAAAAT), FAM123 (Fw:AAGGTTGGTTAGGTGTTTGATG; Rev:TCCAACAACCCTTCCTAAAA), PACRG (Fw:TTAAAGTGTTTATTGGTATTAGGGGT; Rev:AAACCRAACTATAAAACCAACTCC) and C10orf90 (Fw:AAGTTATTGGTATAGAGGGTAGGTAAA; Rev:TTTAAAAACAACCACATAAACCTTAAATA) additioned with Nextera adapter sequences (TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG and GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG). Amplicons were pooled sample-wise and indexed using Illumina Nextera XT Index. Sequencing was performed on an Illumina Miseq Platform with Micro v2 300PE chemistry.