The transfection reagent, Lipofectamine®2000, was selected for optimizing the FIGURE 1. Workflow of PCR product based HEK293 cell transfection (96-well-plate format with Lipofectamine 2000).
‐ HEK293T cells are transiently transfected with proteins of interest using lipofectamine 2000 (invitrogen #11668‐027) and nuclear extracts were prepared using NE‐PER nuclear and cytoplasmic extraction reagents (Thermo Scientific #78833)
Feb 08, 2010 · The DNA-Lipofectamine 2000 complexes were then added to each well containing cells and medium. The vol/wt ratios of Lipofectamine 2000/DNA were: 3/1, 5/1 and 7/1, and 1/1 for Lipofectamine Plus/DNA. Polyethylenimine (PEI) PEI (QBiogene, Morgan Irvine, CA) and plasmid DNA were each diluted with equal volumes of 150 mM NaCl.
Dilute 14 m L of Lipofectamine reagent in 336 m L of RPMI-A in separate eppindorf tube. Mix by tapping bottom of tube on the hood surface. 8. After 15 min incubation in Step 6., add 100 m L of diluted Lipofectamine reagent to each eppindorf tube. 9. Let stand at room temperature for 15 min. 10.
Lipofectamine™ 2000 is supplied in liquid form at a concentration of 1 mg/ml. Take care to maintain a standard seeding protocol between experi-ments because transfection efficiency is sensitive to...
Oct 26, 2020 · Reporter plasmids were transfected at 200 ng per well with lipofectamine 3000 (Invitrogen), following the manufacturer’s protocol. Cells were harvested 16 h post-transfection. Western blot. Cells were lysed with RIPA buffer containing protease inhibitor (EDTA-free, Thermo Fisher Scientific) at 4 °C for 30 min.
Hi, I transfect NIH 3T3 often initially I also used Lipofectamine 2000 and had low efficiency, but then switched to Lipofectamine LTX (slightly more expensive) and it works much better.
The EnGen sgRNA Synthesis Kit, S. pyogenes provides a simple and quick method for transcribing high yields of sgRNA in a single 30 minute reaction, using the supplied reagents and target-specific DNA oligos designed by the user.