RAW264.7細(xì)胞轉(zhuǎn)染操作步驟在已經(jīng)發(fā)表文章中的經(jīng)驗(yàn)分享介紹

石河子大學(xué)動(dòng)物科學(xué)與技術(shù)學(xué)院，在2021年發(fā)表文章到Frontiers in Immunology的學(xué)術(shù)期刊中科研者使用美國zeta life轉(zhuǎn)染試劑，高效率轉(zhuǎn)染質(zhì)粒DNA和siRNA到RAW264.7細(xì)胞，具體轉(zhuǎn)染RAW264.7細(xì)胞方法經(jīng)驗(yàn)如下：（注意：本文轉(zhuǎn)染實(shí)驗(yàn)中使用的細(xì)胞數(shù)量、試劑用量和操作方法不適用于傳統(tǒng)lipo3000轉(zhuǎn)染試劑或類似產(chǎn)品）

All plasmids and siRNAs (20 uM/uL) were transfected using the Advanced DNA  RNA Transfection Reagent™ (ZETA LIFE，USA) according to the manufacturer’s  protocol. Briefly,RAW264.7 cells were planted on the cell culture plate one day in advance, the  cell confluence degree was up to 60 – 80 % at the time of transfection, plasmid or siRNA was directly mixed with transfection reagent according to 1:1 relationship,  then mixed by blowing into a pipette for 10 – 15 times. Following incubation at room  temperature for 10-15 minutes, the complex was added to the cell culture plates, mix 148 gently, and incubated in the CO2 incubator for 24h. All the siRNAs were designed by  Gene Pharma  transfection efficiency was measured using 150 qRT-PCR. 

Zeta Life介紹：

美國 Zeta Life 公司成立于 1989 年，是國際無血清細(xì)胞培養(yǎng)基 DMEM/F12 主要完成人，有三十多年的胎牛血清、無血清細(xì)胞培養(yǎng)基、以及干細(xì)胞、免疫細(xì)胞及腫瘤細(xì)胞等多種不同類型哺乳動(dòng)物細(xì)胞的無血清培養(yǎng)液生產(chǎn)經(jīng)驗(yàn)；也是目前世界胎牛血清、無血清培養(yǎng)基最大的 OEM 供應(yīng)商之一；每批血清均有完整的血源證明文件、獸醫(yī)師檢驗(yàn)證明及品質(zhì)測試報(bào)告；ZETA LIFE 血清在 100 級的無菌間采用全自動(dòng)方式制造生產(chǎn)，具有良好的可再現(xiàn)性及可追溯性。  2018 年 Zeta Life 公司與美國加利福尼亞大學(xué)舊金山校區(qū)聯(lián)合開發(fā)用于哺乳動(dòng)物細(xì)胞、活體動(dòng)物轉(zhuǎn)染的 Advanced DNA RNA 第三代多肽小分子轉(zhuǎn)染試劑，此技術(shù)成為新的全球蛋白功能、免疫細(xì)胞及干細(xì)胞治療、研發(fā)及生產(chǎn)的主要關(guān)鍵技術(shù)之一。