500ml | 250ml | 50ml | Reagent | |
415ml | 207.5ml | 41.5ml | KO DEMEM | |
75ml | 37.5ml | 7.5ml | KO SRM | |
5ml | 2.5ml | 0.5ml | GlutaMax, NEAA, P/S | |
0.5ml | 0.25ml | 0.05ml | ß-mercapto ethanol |
500ml | 250ml | 50ml | Reagent | |
480ml | 240ml | 48ml | DMEM F12 + HEPES | |
5ml | 2.5ml | 0.5ml | GlutaMax, N2, P/S | |
7.5ml | 3.75ml | 0.75ml | 20% Dextrose |
500ml | 250ml | 50ml | Reagent | |
485ml | 242.5ml | 48.5ml | Neurobasal | |
5ml | 2.5ml | 0.5ml | GlutaMax, P/S | |
7.5ml | 3.75ml | 0.75ml | 20% Dextrose | |
10ml | 5ml | 1ml | B27 supplement | |
2.5ml | 1.25ml | 0.25ml | NEAA |
A | B | C | D | E | F | |
Reagent | Vendor | Cat no. | Stock con. | Working conc. | Dilution | |
Neurobasal medium | Thermofisher Scientific | #21103049 | n.a. | n.a. | n.a. | |
Knock DMEM | Gibco | #10829-018 | n.a. | n.a. | n.a. | |
KnockOut Serum Replacement | Invitrogen | 10828-028 | n.a. | n.a. | n.a. | |
MEM non-essential amino acids | Invitrogen | 11140-050 | n.a. | n.a. | n.a. | |
Beta-mercaptoethanol | Invitrogen | 21985-023 | n.a. | n.a. | n.a. | |
GlutaMax | Invitrogen | 35050061 | n.a. | n.a. | n.a. | |
DMEM F12 HEPES | Gibco | 11330057 | n.a. | n.a. | n.a. | |
N2 supplement | Invitrogen | 17502048 | n.a. | n.a. | n.a. | |
20% Dextrose | 20% | n.a. | n.a. | |||
Penicillin/Streptomycin | Invitrogen | 15140122 | 100x | 1x | 1:100 | |
Trehalose | Sigma | #T9531-25G | n.a. | n.a. | n.a. | |
DMSO | Sigma | #D8418-250M | n.a. | n.a. | n.a. | |
mTeSR plus | Stem cell Technologies | # 100-0276 | n.a. | n.a. | n.a. | |
Matrigel (Differentiation) | Corning | 354234 | n.a. | n.a. | n.a. | |
Dnase | NEB | #M030 | n.a. | n.a. | n.a. | |
BDNF | Peprotech | #450-02 | 10µg/ml | 10ng/ml | ||
GDNF | Peprotech | #450-10 | 10µg/ml | 10ng/ml | ||
CNTF | Peprotech | # 450-13 | 10µg/ml | 10ng/ml | ||
DAPT | Fisher Scientific | #26-341-0 | 10mM | 10µM | ||
Doxycyclin Hyclate | Sigma | D9891-5g | 2mg/ml | 2µg/ml | ||
Puromycin | Life technology | A11138-03 | 10mg/ml | |||
Accutase | Gibco | #A11105 | ||||
Y27632 ROCK Inh. | Stem cell technologies | #72304 | 10mM | 10µM | ||
Cell strainer (40µm) | Falcon/Corning | #352340 | n.a. | n.a. | n.a. | |
B27 Supplement | Invitrogen | 17504044 | n.a. | n.a. | n.a. |