title: MinION sequencing; Nanopore sequencing of UNC13A from SHSY5Y and patient RNA
design: Nanopore sequencing of UNC13A from SHSY5Y and patient RNA
library_construction_protocol: SH-SY5Y cells were transduced with SmartVector lentivirus (V3IHSHEG_6494503) containing a doxycycline-inducible shRNA cassette for TDP-43. Transduced cells were selected with puromycin (1 g/mL) for one week. RNA was extracted with Trizol and column cleanup.  RNA was extracted with Trizol and column cleanup Following reverse transcription with specific primers, nested PCR (keeping cycle number low) was used to obtain pure amplicons with Nanopore-compatible overhangs. Nanopore barcodes were then added and Nanopore adapters were added via rapid adapter addition.