Single-cell genomic sequencing of PC9 cell line
The single-cell sequencing protocol was adapted from (Baslan et al. Nat. Protoc. 2012). Single-cell genomic library prepared from single cells flow-sorted into lysis buffer. Genomic DNA fragmented enzymatically with NlaIII. Both ends of genomic fragments were tagged using oligonucleotides containing a cell-barcode, a universal primer, and several random nucleotides (varietal tags). Cell barcodes allow for multiplexing of the samples and the varietal tags allow for counting of unique initial DNA fragments. After amplification with a universal primer, fragment size selection was performed with Agencourt AMPure XP beads (Beckman Coulter; A63881) to obtain fragments appropriate for sequencing. Barcoded sequencing adaptors were ligated to the ends of the fragments to allow for further multiplexing and to prepare the fragments for next generation sequencing. Next generation sequencing was performed on an Illumina MiSeq