如何高效率转染HCT-116人结直肠癌细胞?
发布网友
发布时间：2022-04-29 05:01
共2个回答
热心网友
时间：2022-06-19 00:55
结肠癌细胞很少有转染试剂能转的了，我们原来都是用病毒感染，前段时间看到西南大学的一篇文章写的是Zetalife有，名字Advanced DNA/RNA Transfection Reagent转染试剂，我们后面用了转染效率很高，分享给大家。
转染条件
A、HCT-116, SW480, HT-29人结直肠癌细胞
B、AURKA质粒
C、转染细胞融合度50%
D、96孔板每孔使用0.5μg质粒DNA
6孔每孔使用10 ug质粒DNA
(注意：本实验中用到的细胞密度、转染质粒DNA用量不适用于LIPO3000/2000)
发表文章部分内容
Overexpression of AURKA Plasmid for AURKA overexpression was obtained from VectorBuilder. Details about plasmid containing the AURKA gene can be found at https://www.vectorbuilder.cn/vector/VB190802-1063ncc. html. colon cancer cells (HCT-116, SW480, HT-29) were seeded at 50% confluency for 24 h in six-well and 96-well plates. Plasmid (10 ug per-well for six-well and 0.5 μg per-well for 96 plates) and the Advanced DNA RNA transfection（zeta life,USA) reagent were mixed in equal proportionsand added to the cell culture medium for 6 h. DOX (1 μg/ml) was added to ince the expression of AURKA. After addition of DOX and PAL for 36 h, cell viability was measured by MTT and apoptosis was detected by。
PAL exerts anti-colon cancer effects by targeting AURKA. (A) PAL promotes apoptosis in colon cancer cells (HCT-116, SW480) in a dose-dependent manner. (B) PAL promotes G2/M phase arrest in colon cancer cells (HCT-116, SW480) in a dose-dependent manner. (C) The expression of AURKA after DOX treatment for 24 h. (D) Cell density after PAL treatment for 24 h in the cells (HCT-116, SW480) transfected with AURKA overexpression plasmids. DOX is an incer of AURKA
expression in plasmids. (E) Effect of PAL detected by MTT assay on the proliferation ability of colon cancer cells transfected with AURKA overexpression plasmids. (F) Effects of PAL detected by.
热心网友
时间：2022-06-19 00:56
