title: MinION sequencing; Targeted nanopore resequencing and methylation analysis of LINE-1 retrotransposons
design: Targeted nanopore resequencing and methylation analysis of LINE-1 retrotransposons
library_construction_protocol: Cells were grown and collected from culture dishes. Genomic DNA was extracted using the Monarch Genomic DNA Purification kit. 125 sgRNAs targeting the region downstream of L1HS elements (one unique sgRNA per locus) were synthesized using the EnGen sgRNA Synthesis kit (New England Biolabs). Targeted librairy preparation follows these steps: assembly of Cas9 ribonucleoprotein particles (RNP), dephosphorylation of genomic DNA, incubation of RNP assemble with genomic DNA, library preparation and Oxford Nanopore sequencing. With this design, sequencing starts at the sgRNA binding site in the region downstream of L1 and progresses towards the target L1 in the antisense direction