Alphavirus-Based Vectors - Viral Vector Approaches in Neurobiology and Brain Diseases - page 96

13. Phosphate-buffered saline (PBS) (Gibco BRL)
14. Trypsin-ethylenediaminetetraacetic acid (EDTA) (0.25 %
Trypsin, 1 mM EDTA × 4 Na) (Gibco BRL)
15. Microcentrifuge, 1.5 mL Microcentrifuge tubes (Eppendorf)
16. Heating blocks and water baths (Eppendorf/Julabo)
17. Sterile electroporation cuvettes, 0.2 and 0.4 cm (BioRad or BTX)
18. Electroporator (BioRad Gene Pulser)
19. Tissue culture fl asks (T25, T75, and T175) (Nunc Brand
Products)
20. Microwell plates (6-, 12-, and 24-well plates) (Costar)
21. Falcon tubes (15 and 50 mL) (Becton Dickinson)
22. Plastic syringes (1, 10, and 50 mL) (Becton Dickinson)
23. Sterile 0.22 m fi lters (Millipore)

24. Glass capillaries (e.g., Clark Electromedical Instruments,
Pangbourne, UK)
25. Electrode puller
26. Electrode holder (airtight)
27. Autoclavable electrode holder (e.g., metal bin with foam)
28.

μ

Micromanipulator (e.g., Narishige)
29. Metal plate containing a base for a 35-mm Petri dish
30. 35 mm Petri dishes (Costar)
31. 3-Way valve
32. Plastic tubing (inner dimension 1 mm, outer dimension 3 mm)
33. Dissection microscope
34. Autoclaved microloader pipet tips (Eppendorf)
35. Forceps, small scissors
36. Cutting medium (Roller-tube culture medium, 10 mM MgCl 2 ,
0.5

M tetrodotoxin, e.g., Latoxan)
37. Hippocampal slice cultures (e.g., roller-tube type)
38. Burner
39. MicroSpin™ S-200 HR Columns (Amersham)
40. Dulbecco's modifi ed F-12 medium (Gibco BRL)
41. Iscove's modifi ed Dulbecco's medium (Gibco BRL)
42. Opti-MEM I reduced-serum medium (Gibco BRL)
43. Neurobasal medium (Invitrogen)
44. X-gal stock solutions: 50 mM K ferricyanide (+4 °C); 50 mM K
ferrocyanide (+4 °C), 1 M MgCl (room temperature), 2 %
X-gal in DMF or DMSO (−20 °C)
45. X-gal staining solution: 1× PBS (1/10 of stock), 5 mM K
ferricyanide (1/10 of stock), 5 mM K ferrocyanide (1/10 of

μ