如何得到稳定转染的克隆
关键词: 来源: 互联网
问:
各位园友,本人今天刚刚做了转染,用的24孔板,质粒是带有荧光的(含neo基因的载体),如果转进去了,下一步就要进行筛选?但是不知道具体该怎么做才能得到稳定转染的细胞?
答:
最好先做以下预实验,摸一下G418的浓度范围,以未转染的细胞做对照,当未转染细胞完全杀死作为最佳筛选浓度。在选择压力下待存活的细胞逐渐形成小克隆后可以传代,随后可以逐步降低G418浓度至原使用浓度的一半作为维持压力。这样多代选择出来的细胞就是阳性克隆,也就是稳定细胞系了。
推荐方法
- Evaluation of Chemosensitivity of Micrometastases with Green Fluorescent Protein Gene-Tagged Tumor Models in Mice
- DNA Methylation Analysis in Human Cancer
- A Simple Method for Profiling miRNA Expression
- The Application of Differential Display as a Gene Profiling Tool
- The Effects of Butyrate and the Role of c-myc in N.1 Ovarian Carcinoma Cells Determined by Northern Blotting
- Analysis of Genome-Wide DNA Methylation Profiles by BeadChip Technology
- Photodynamic Therapy in Lung Cancer: A Review
- pIRES2-eGFP转染细胞的经验
- 细胞凋亡后beta-actin会降解吗?
- CHO细胞表达系统