GSM5216898: PBAT_bulk_DAC_treated [PBAT-DAC_S51_L001]; Homo sapiens; OTHER
We utilised the G&T-seq protocol to separate genomic DNA and RNA from the single-cell samples [Macaulay, I.C., et al., G&T-seq: parallel sequencing of single-cell genomes and transcriptomes. Nature Methods, 2015.]. Genomic DNA from each cell was purified and bisulphite conversion was performed as described [Angermueller, C., et al., Parallel single-cell sequencing links transcriptional and epigenetic heterogeneity. Nature methods, 2016], with minor modifications. A post-bisulphite adaptor tagging (PBAT) approach protocol [Miura, F., et al., Amplification-free whole-genome bisulfite sequencing by post-bisulfite adaptor tagging. Nucleic Acids Research, 2012] was used to prepare genome-wide sequencing libraries from bulk samples. Libraries were prepared as described [Rulands, S., et al., Genome-Scale Oscillations in DNA Methylation during Exit from Pluripotency. Cell Systems, 2018]. The only alteration from this protocol is the use of a new 6NR adaptor 2 oligo (5'-CAGACGTGTGCTCTTCCGATCTNNNNNN-3') that is suitable for use with NEBNext dual index oligo kits.