R_pacifica_Nanopore
A total of 600ng of purified gDNA from the sample was end-repaired (NEBnext ultra II end repair kit, New England Biolabs, MA, USA), cleaned up with 1x AMPure beads (Beckmann Coulter, USA). Native barcode ligation was performed with NEB blunt/ TA ligase (New England Biolabs, MA, USA) using EXP-NBD114 (ONT) and cleaned with 1xAmPure beads.Qubit quantified barcode ligated DNA sample was Adapter ligated for 15 minutes using NEBNext Quick Ligation Module (New England Biolabs, MA, USA). Library was cleaned up using 0.6X AmPure beads (Beckmann Coulter, USA) and finally sequencing library was eluted in 15 l of elution buffer and used for sequencing.Sequencing was performed on GridION X5 (Oxford nanopore Technologies, Oxford, UK) using SpotON flow cell R9.4 (FLO-MIN106) in a 48 hrs sequencing protocol