Target bisulfite sequencing of human: whole blood of relapsed/refractory B cell lymphoma patient after CD19.CAR T cell therapy
A two-step PCR approach on bisulfite-converted DNA was adopted to generate target bisulfite sequencing libraries. A multiplex PCR was performed combining bisulfite-specific primers for APC2 (Fw:GGTTYGGGTTTTTGGTGA; Rev:CCAACCCAACCTCATTACTA), FAM123 (Fw:AAGGTTGGTTAGGTGTTTGATG; Rev:TCCAACAACCCTTCCTAAAA), PACRG (Fw:TTAAAGTGTTTATTGGTATTAGGGGT; Rev:AAACCRAACTATAAAACCAACTCC), cg02619656 (Fw:ATTTGATTTTGTYGTAGTGGA; Rev:AAAATCCCRACCTCTACTTAA), cg09787504 (Fw:TATATGTGTGTAYGTTGAATAAAAT; Rev:TCCATTTCATATCAATACTAATATT) additioned with Nextera adapter sequences (TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG and GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG). PCR product was indexed using Illumina Nextera XT Index. Sequencing was performed on an Illumina Miseq Platform with Micro v2 300PE chemistry.