Supplementary Fig. 4: Sequencing library preparation.
From: High-throughput single-cell ChIP-seq identifies heterogeneity of chromatin states in breast cancer
(a) Enriched barcoded nucleosomes were linearly amplified by in vitro transcription. The amplified RNAs were reverse transcribed into cDNA by random priming, appending a reverse complement of Illumina Read #1 sequencing primer. The cDNAs were amplified by PCR, appending an Illumina P7 and P5 sequences. (b) Schematic of the final sequencing product with size in bp of each element constituting the sequence. (c) Electropherogram showing the size distribution of the final sequencing library post agarose gel purification. The smear ranges from 300 bp to 700 bp and corresponds to barcoded nucleosomes (profile obtained by Tapestation [Agilent]). (d) Single-cell ChIP-seq libraries were sequenced as follows: 50 bp were assigned to read the nucleosomal sequence and 100 bp were assigned to read the barcode.
