[求救] Two-stage Cell transformation
想請問一下有人做過two stage cell transformation的實驗嗎
我是用MNNG以及TPA處理當作positive control
之後做soft agar assay看細胞有沒有trasnformation
可是做出來都沒有colony
不知道哪裡出了問題
以下是我的protocol
1. Treat 2ug/ml of MNNG for 72hr
2. Remove MNNG containing media and culture cells for 3 days in normal
conditioning media without additives.(我是用serum free medium)
3. Culture cells for 2 more weeks in conditioning media containing
100ng/ml TPA.
4. Following TPA treatment, culture cells one more week in normal conditional
media, without assitives.
5. Continue with soft agar assay.
我的soft agar base layer是0.5%
top layer是0.33%
1000 cells/well(6 well plate)
cell line是mouse embryo liver cell
先謝謝大家的回答
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