qiagen PolyFect Transfection Reagent DNA轉(zhuǎn)染試劑
qiagen PolyFect Transfection Reagent DNA轉(zhuǎn)染試劑
PolyFect Transfection Reagent采用的活化樹狀多聚物,從多聚物中心向外輻射出帶正電荷的氨基基團(tuán)與DNA 分子結(jié)合,為下列細(xì)胞株特別優(yōu)化:COS-7、NIH/3T3、HeLa、293、CHO。
qiagen PolyFect Transfection Reagent DNA轉(zhuǎn)染試劑
olyFect Reagent用于HeLa細(xì)胞轉(zhuǎn)染。
圖中顯示了HeLa細(xì)胞中[A]β-半乳糖苷酶和[B]綠色熒光蛋白(GFP)的表達(dá)。依據(jù)PolyFect Transfection Reagent與HeLa細(xì)胞轉(zhuǎn)染實(shí)驗(yàn)方案,在6孔孔板中將β-半乳糖苷酶和GFP報(bào)告質(zhì)粒共轉(zhuǎn)染入細(xì)胞中。轉(zhuǎn)染兩天后,通過X-gal染色或熒光顯微鏡觀察基因表達(dá)情況。
在CHO和293細(xì)胞中具有高轉(zhuǎn)染效率。
對比PolyFect Reagent和兩個(gè)脂質(zhì)試劑的轉(zhuǎn)染效率。給培養(yǎng)在6孔孔板中的細(xì)胞轉(zhuǎn)染β-半乳糖苷酶報(bào)告質(zhì)粒。使用PolyFect Reagent轉(zhuǎn)染時(shí)依據(jù)相應(yīng)實(shí)驗(yàn)方案進(jìn)行。Reagents L2 和F的轉(zhuǎn)染依據(jù)優(yōu)化的實(shí)驗(yàn)方案。對于CHO細(xì)胞:10 µl Reagent L2轉(zhuǎn)染3 µg DNA,15 µl Reagent F轉(zhuǎn)染2 µg DNA;對于293細(xì)胞,10 µl Reagent L2轉(zhuǎn)染2 µg DNA,7 µl Reagent F轉(zhuǎn)染3 µg DNA。所有轉(zhuǎn)染均一式三份進(jìn)行。圖中展示了293細(xì)胞的兩次獨(dú)立轉(zhuǎn)染實(shí)驗(yàn)。
PolyFect轉(zhuǎn)染效率。
轉(zhuǎn)染流程簡單、快速。將PolyFect Reagent加入DNA溶液中,混勻、孵育5–10分鐘,加入培養(yǎng)及(可含有血清和抗生素),將PolyFect-DNA加入細(xì)胞中。孵育細(xì)胞,進(jìn)行基因表達(dá)。
Cat No./ID: 301105 PolyFect Transfection Reagent (1 ml) For 25–65 transfections in 60 mm dishes or 50–100 transfections in 6-well plates. |
Cat No./ID: 301107 PolyFect Transfection Reagent (4 x 1 ml) For 100–260 transfections in 60 mm dishes or 200–400 transfections in 6-well plates. |
Cat No./ID: 301108 PolyFect Transfection Reagent (100 ml) For 2500–6500 transfections in 60 mm dishes or 5000–10,000 transfections in 6-well plates. |