Gene Therapy for Epilepsies - Gene Delivery and Therapy for Neurological Disorders - page 245

40 % iodixanol

50 ml 10× PBS.
0.5 ml 1 M MgCl 2 .
0.5 ml 2.5 M KCl.
333 ml iodixanol.
Make up to 500 ml with distilled water.
Filter sterilize through a 0.22 μm nitrocellulose filter.
Store at 4 °C.
54 % iodixanol
50 ml 10× PBS.
0.5 ml 1 M MgCl 2 .
0.5 ml 2.5 M KCl.
449 ml iodixanol.
Filter sterilize through a 0.22 μm nitrocellulose filter.
Store at 4 °C.

2×PBS/1 mM MgCl 2
100 ml 10× PBS.
0.5 ml 1 M MgCl 2 .
Make up to 500 ml with distilled water.

15-cm tissue culture dishes
50-ml disposable polypropylene conical tubes
Cell scrapers
Refrigerated high-speed centrifuge
Dry ice/ethanol bath
37 °C water bath
OptiSeal ultracentrifuge tube
Ultracentrifuge with fixed angle rotor (e.g., we use Beckman
Coulter Optima L-100 XP with 70 Ti Beckman Coulter rotor)
Clamp stand
20-G needle
10-ml syringe
Amicon 100 K MWCO concentrator
Centrex microfilter
Screw cap microtubes for vector storage (e.g., VWR 16466-050)

2.2 Genomic Titering
of AAV Vectors

Plasmid standards

PCR buffer
500 mM KCl
100 mM Tris, pH 8.0
50 mM MgCl 2