title: TPK2.0 preparation of E14 mouse embryonic stem cell line genomic DNA
design: Bulk gDNA was extracted from E14 mouse embryonic stem cell lines at Passage 33 via phenol:chloroform:isoamyl alcohol extraction. Approximately 2.5g purified DNA was fragmented to 6-8 kb using a g-TUBE (PN: 520079, Covaris) with an Eppendorf 5424 rotor spun at 7,000 rpm for 6 passes. Sheared DNA was used as input for the PacBio SMRTbell Express Template Prep Kit 2.0 protocol (TPK2.0). Briefly, the TPK2.0 protocol consists of removal of single stranded overhangs, DNA damage (PreCR) repair, end-repair, A-tailing, barcoded SMRTbell adapter ligation, and exo digestion followed by 1X AMPure PB bead cleanup.