Table 1.
Materials (reagents, supplies, and biologics)
| Material | Supplier | Cat. no. | Specific handling | Storage |
|---|---|---|---|---|
| Reagents | ||||
| DMEM + GlutaMAX | Invitrogen/Gibco | 61965 | 4°C | |
| FBS | Biochrom AG | S 0115 | −20°C | |
| Penicillin–streptomycin | Invitrogen/Gibco | Various | −20°C | |
| Liquid nitrogen | Various | |||
| TurboFect transfection reagent | Thermo Scientific | R0531 | 4°C | |
| PEG-6000 | Sigma–Aldrich | 81260 | RT | |
| Sodium hydroxide | Sigma–Aldrich | S7653 | Corrosive | RT |
| Hydrochloric acid 37% | Sigma–Aldrich | 320331 | Corrosive | RT |
| Sodium chloride | Sigma–Aldrich | S8045 | RT | |
| PEI | Polyscience | 23966 | RT | |
| PFA | Electron Microscopy Science | 15710 | RT | |
| Hoechst 33258 | ThermoFisher Scientific | H1398 | 4°C | |
| PTA | Serva | 32757.01 | RT | |
| Pioloform BM18 | Plano GmbH | R1275B | ||
| Quant-IT™ DNA assay kit | ThermoFisher Scientific | Q33210 | 4°C | |
| Quant-IT protein assay kit | ThermoFisher Scientific | Q33120 | 4°C | |
| ddPCR Supermix for probes | Bio-Rad | 186310 | −20°C | |
| Supplies | ||||
| Bottle top filter 0.2 μm | VWR | 514-0340 | RT | |
| Syringe filters 0.8, 0.65, 0.45, and 0.2 μm | Sartorius Stedim Biotech | Various | RT | |
| Regenerated cellulose membranes 1.0 μm | Cytiva | 10410014 | RT | |
| Stainless steel filter holder 25 mm | Merck Millipore | XX3002500 | RT | |
| Superdex 200 Increase 10/300 GL column | Cytiva | 17517501 | RT | |
| Rubber O-ring | Cytiva | 18-1029-60 | RT | |
| Hole puncher (25 mm) for regenerated cellulose membranes | Various | DIN 7200 A | RT | |
| 1.5–2.0 mL centrifuge tubes | Various | RT | ||
| 15 and 50 mL centrifuge tubes | Various | RT | ||
| Disposable syringes, 10–20 mL | Various | RT | ||
| Cell lifter, high-density polyethylene | Corning | 3008 | RT | |
| 15 cm dishes | Various | RT | ||
| 6-well plates | Various | RT | ||
| Flasks 75 cm2 (T75) | Various | RT | ||
| 96-well flat bottom assay plate, black polystyrene | Corning | 3915 | RT | |
| 250 mL centrifuge tubes | Sigma-Aldrich | CLS430776 | RT | |
| Whatman grade 1 filter paper | Cytiva | 1001-110 | RT | |
| Twin.tec 96-well PCR plate | Eppendorf AG | Various | ||
| Dialysis membranes, 100 or 300 kDa cutoff | Spectrum | 131414 or 131450 | 4°C | |
| 300 mesh copper grids | Plano GmbH | G2430C | RT | |
| Biologics | ||||
| HEK293T cells | ATCC | CRL-3216 | −80°C | |
| AAV helper plasmid (encoding the cap gene of AAV1, AAV2, or AAV6) | Ref.21 | −20°C | ||
| Benzonase® or Denarase® endonuclease | Merck Millipore or c-LEcta | −20°C | ||
AAV, adeno-associated virus; ddPCR, Droplet Digital™ polymerase chain reaction; DMEM, Dulbecco's modified Eagle medium; FBS, fetal bovine serum; PEG, polyethylene glycol; PEI, polyethylenimine; PFA, paraformaldehyde; PTA, phosphotungstic acid; RT, room temperature.