Lentiviral Vectors for Gene Delivery to the Nervous System - Gene Delivery and Therapy for Neurological Disorders - page 38

Table 3
(continued)

Volume Components Final concentration

TVM2 media (pH 7.90)
326.8 ml

Cell culture grade H 2 O (SIGMA # W3500)

5.52 g DMEM-H powder (SIGMA D5648-SL)

10 ml 1 M HEPES (SIGMA H3537)

40 ml Newborn Calf Serum (SIGMA) 10 %

19.72 ml 7.5 % sodium bicarbonate solution (SIGMA, S8761)

4 ml 10× Pen-Strep (Penicillin 10,000 U/ml; Streptomycin 10,000 ʼ g/ 1 %

ml, SIGMA)

20 % Sucrose solution
80 ml

Cell culture grade H 2 O (SIGMA # W3500)

20 g Sucrose powder (SIGMA # 84097) 20 %

2 ml 5 M NaCl (SIGMA, # S5886)

2 ml 1 M HEPES solution pH 7.4 (GIBCO, #15630-049)

200 ʼ l 0.5 M EDTA solution

4 % Paraformaldehyde
4 g Paraformaldehyde powder (SIGMA # P6148) 4 %

ʼ

100 l 10 M NaOH

10 ml PBS (SIGMA, # D8537)

85 ml Cell culture grade H 2 O (SIGMA # W3500)

1 M CaCl 2
11.1 g

CaCl 2 powder (SIGMA # 22350-6) 1 M

100 ml Cell culture grade H 2 O (SIGMA # W3500)

Table 4
List of reagents and consumables

Reagents and consumables Sources and catalog numbers

Ampicillin sodium salt SIGMA, A9518-5G

Cryotube 1.0 ml Nunc, 366656

DAPI SIGMA

1 kb DNA ladder NEB, #N3232

EndoFree plasmid mega kit [ 5 ] QIAGEN, 12381

50 ml Falcon tubes VWR, 525-0156

HEK 293T cell line ATCC (CRL-11268™) [HEK293T/17] Rockefeller Univ.

Lenti-X qRT-PCR titration kit Clontech, Cat. No. 631235

Newborn calf serum (heat inactivated) SIGMA, #N4762-500ML

OCT Surgipath FSC22, Leica Microsystems, Wetzlar, Germany

(continued)