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PromoCell Airway Epithelial Cell Growth Medium is a serum-free medium optimized for the in vitro cultivation of epithelial cells from large air passages.
Although all PromoCell media are optimized for use with primary human cells, we have received feedback from customers that this particular medium can also be used for equine, porcine, murine, bat and rat airway epithelial cells.
To view a list of references where this medium was used with other species/cell types please click here.
Components:
- Growth Medium (Ready-to-use): Includes Basal Medium and SupplementMix
- Growth Medium Kit: Includes Basal Medium and SupplementPack
- SupplementMix: Contains all media supplements premixed in one vial
- SupplementPack: Contains all media supplements as individual vials
Media supplement | Final supplement concentration |
---|---|
Bovine Pituitary Extract | 0.004 ml / ml |
Epidermal Growth Factor (recombinant human) | 10 ng / ml |
Insulin (recombinant human) | 5 μg / ml |
Hydrocortisone | 0.5 μg / ml |
Epinephrine | 0.5 μg / ml |
Triiodo-L-thyronine | 6.7 ng / ml |
Transferrin (recombinant human) | 10 μg / ml |
Retinoic Acid | 0.1 ng / ml |
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由于质粒的不兼容性,拥有同种复制子的质粒不能在同一细胞内稳定共存,经过几代的复制,会质粒丢失,所以并不是任何两个或两个以上质粒都可以在同一细胞内稳定存在,但是可以同时进入.
稳定转染的细胞株,就是转染后质粒可以稳定整合到基因组上不会因细胞分裂而丢失。区别于质粒瞬时转染不能长时间保留质粒在细胞里。
如果经费充足的话可以找公司包装病毒,如武汉的普健可以提供各种载体的构建及细胞株构建的技术服务。
先提取RNA,反转录成cDNA,然后根据目的基因设计PCR引物,通过半定量RT-PCR确定目的基因表达.
正因为检测的是mRNA,所以要先反转录成cDNA才能PCR.
2.将构建完成的载体与慢病毒包装质粒混合,共转染靶细胞
3.收集病毒液
4.用病毒液感染靶细胞
5.用载体上带的抗生素进行筛选,如果没有,可以用无限稀释法
6.获得稳转株
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