石河子大学动物科学与技术学院,在2021年发表文章到Frontiers in Immunology的学术期刊中科研者使用美国zeta life转染试剂,高效率转染质粒DNA和siRNA到RAW264.7细胞,
具体转染RAW264.7细胞方法经验如下:(注意:本文转染实验中使用的细胞数量、试剂用量和操作方法不适用于传统lipo3000转染试剂或类似产品)
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介绍:
美国 Zeta Life 公司成立于 1989 年,是国际无血清细胞培养基 DMEM/F12 主要完成人,有三十多年的胎牛血清、无血清细胞培养基、以及干细胞、免疫细胞及肿瘤细胞等多种不同类型哺乳动物细胞的无血清培养液生产经验;也是目前世界胎牛血清、无血清培养基最大的 OEM 供应商之一;每批血清均有完整的血源证明文件、兽医师检验证明及品质测试报告;ZETA LIFE 血清在 100 级的无菌间采用全自动方式制造生产,具有良好的可再现性及可追溯性。 2018 年 Zeta Life 公司与美国加利福尼亚大学旧金山校区联合开发用于哺乳动物细胞、活体动物转染的 Advanced DNA RNA 第三代多肽小分子转染试剂,此技术成为新的全球蛋白功能、免疫细胞及干细胞治疗、研发及生产的主要关键技术之一。