Target bisulfite sequencing of human: Post-mortem Brain - Frontal Cortex (BA9)
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 cg23661000 (Fw:TTTAGTGTTAGAATTGAAAGAGTAGA; Rev:TTAACCTTAACTATATCTAACAAAAA), cg09787504 (Fw:TATATGTGTGTAYGTTGAATAAAAT; Rev:TCCATTTCATATCAATACTAATATT) and cg02619656 (Fw:ATTTGATTTTGTYGTAGTGGA; Rev:AAAATCCCRACCTCTACTTAA) (previously published in Lubotzky et al, 2022/10.7554/eLife.76391) 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.