Biology Reference
In-Depth Information
10 cm Petri dishes (Falcon)
T150 cm 2 fl ask (Corning)
Cellstack (Corning)
Production
0.5 M CaCl 2 (Sigma)
2 × HEPES-buffered saline (Sigma)
Stericup fi lter (0.45
m pores; Millipore)
Ultracentrifuge equipment (rotors 32Ti and SW55) (Beckman)
1 % Bovine serum albumin (BSA) (Sigma)
Low-retention tubes (Axigen)
μ
2.3 LV Titration
HIV-1 p24 ELISA kit (RETROtek, Gentaur)
DNase I, amplifi cation grade (Invitrogen)
RNase A (Invitrogen)
RNasin (Invitrogen)
Ethylenediaminetetraacetic acid (EDTA) (Sigma)
Express Superscript Mix (Invitrogen)
Express SyberGreenER Qpcr Super Mix Univ (Invitrogen)
PureLink viral RNA/DNA kit (Invitrogen)
DNase- and RNase-free water (Gibco)
2.3.1 p24
2.3.2 qPCR
24-well plate (Sigma)
37 % Formaldehyde 37 % (Sigma)
5 ml Tube for fl ow cytometry (Falcon)
2.3.3 FACS
2.4 Primary Striatal
Cultures
PBS: Dilute PB to 0.1 M and add 9 g/l NaCl (Gibco)
Poly- D -lysine (30-70 kDa; Sigma): 5 mg in 100 ml sterile water
1 M HEPES (Sigma) (100×)
100 mM sodium pyruvate (Sigma) (100×)
HBSS without Ca 2+ and Mg 2+ (Hank Balance Salt Solution)
(Gibco)
HBSSm: 1× HEPES, 1× sodium pyruvate in HBSS
6-well plates, 24-well plates, large Petri dishes (Falcon), pair of
forceps for dissection, microscissors
Isofl urane (Abbott), pentobarbital (Sanofi )
DNase I (3 mg/ml) (Sigma): use at 300
2.4.1 Cultures of Primary
Neurons and Astrocytes
g/ml in HBSSm
Neuronal culture medium: Neurobasal (Gibco) supplemented
with 2 % B27 (Invitrogen)
μ
Search WWH ::




Custom Search