Adeno-Associated Virus-Based Vectors - Viral Vector Approaches in Neurobiology and Brain Diseases - page 43

in the presence of adenovirus. To determine the level of
contamination, the DNA membranes are therefore hybrid-
ized to a specific rep-cap probe. Alternatively, quantitative
real-time PCR using specific primers for rep or cap can be
directly performed on crude lysate.

4 Notes

Institutional guidelines should be followed for the safe handling of
infectious agents and radioactive materials.
All virus purification and titration steps should be conducted inside
a level II biosafety cabinet. Decontamination and disposal of
all material that come in contact with viruses should be inacti-
vate with virucidal agents (e.g., virkon).
Radioactive materials should be stored and handled only in desig-
nated area and suitable shielding should be used.

5 Annexes

15 % iodixanol/1 M NaCl
Add 45 ml of Optiprep (iodixanol), 36 ml of 5 M NaCl, 36 ml of
5× TD, and 63 ml of ddH 2 O. Filter sterilize and aliquot in
appropriate sizes. Store aliquots protected from light at room
temperature.
25 % iodixanol/phenol red
Add 100 ml of Optiprep (iodixanol), 48 ml of 5× TD, 92 ml of
ddH 2 O, and 600 μl of phenol. Filter sterilize and aliquot in
appropriate sizes. Store aliquots protected from light at room
temperature.
40 % iodixanol
Add 136 ml of Optiprep (iodixanol), 40 ml of 5× TD, and 24 ml
of ddH 2 O. Filter sterilize and aliquot in appropriate sizes.
Store aliquots protected from light at room temperature.
60 % iodixanol/phenol red
Add 200 ml of Optiprep (iodixanol) and 500 μl of phenol red.
Filter sterilize and aliquot in appropriate sizes. Store aliquots
protected from light at room temperature.
6× Laemmli buffer
Add 1.2 g SDS, 6 mg bromophenol blue, 4.7 ml glycerol, 1.2 ml
Tris 0.5 M, pH 6.8, and 2.1 ml ddH 2 O. Stir until it dissolves.
Add 0.93 g DTT. Stir until it is completely dissolved. Aliquot
and store at −20 °C.

5.1 Preparation of
Buffers and Solutions