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文献上说得很简略，找了很久终于找到应其龙原始的配方。引至Embryonic stem cell protocol这本书，希望对大家有帮助。
1 ?8 r' B$ a% H K1. N2 100X stock solution. For 10 mL, mix 1 mL insulin (2.5 mg/mL final concentration) with 1 mL apo-transferrin (10 mg/mL final concentration), 0.67 mL BSA (5 mg/mL final concentration), 33 μL progesterone (2 μg/mL final concentration), 100 μL putrescine (1.6mg/mL final concentration), 10 μL sodium selenite (3 mM final concentration), and 7.187mL DMEM/F12. Store at 4°C and use within 1 mo.
5 S) _# Q) [, A% x2. DMEM/F12-N2 medium: to 100 mL DMEM/F12, add 1 mL N2 100X stock solution. The final concentration of each component of N2 in the DMEM/F12 medium is 25 mg/mL insulin; 100 mg/mL apo-transferrin; 6 ng/mL progesterone; 16 mg/mL putrescine; 3 nM sodium selenite; 50 mg/mL BSA. Store at 4°C and use within 1 mo.1 ^# Y2 a7 X. z+ a
3. Neurolbasal/B27 medium: to 100 mL Neurolbasal medium, add 2 mL B27 and 0.5–1 mL 200 mM L-glutamine. Store at 4°C and use within 1 mo.
/ X8 h. l9 n: d7 ^- Y, r4 ?! O4. N2B27 medium: mix DMEM/F12-N2 medium with Neurolbasal/B27 medium in the ratio of 1:1. Add β-mercaptoethanol to a final concentration of 0.1 mM from the 0.1 M stock. Store at 4°C and use within 1 mo. |
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发表于 2015-1-30 10:19
