• Jun 20, 2020 · Day 2: Transfect cells with a plasmid expressing the peptide M9M (Cansizoglu et al., 2007) using lipofectamine 3000 (Thermo Fisher Scientific). Note: Express M9M in fusion with a fluorescent protein (e.g., Red fluorescent protein) to identify transfected cells. Change medium 4 h after transfection to reduce toxicity. Incubate cells overnight.
  • Lipofectamine 3000 is the way to go! 😁 jk I haven’t tried Calcium phosphate so can’t comment on that.
  • Transfections were conducted in a 24-well plate with 300 ng pX330, 400 ng pEF-BFP-Puro R, 50 ng sgRNA vector, 1.5 μl Lipofectamine 3000 (ThermoFisher Scientific) and 1 μl P3000 transfection reagent. Cells were passaged at 72 h post-transfection into a single well of a 6-well plate and selected with 0.5 μg/ml puromycin for 2 weeks.
  • I use lipofectamine 3000 to transfect Hek 293 cells in 6 wells plates. For the last transfection, I transfected at 60% confluency. Also, I used 2500 ng of DNA per well, 5µl of P3000 per DNA, 5µl ...
  • Results for the optimization of lipofection conditions for HMC-1.2 cells using Lipofectamine 3000 are available in Supplementary TableS1.Thephysicalproperties(size,polydispersity,andDNAencap-sulation efficiency) of the various plasmid-containing LNP formula-tions tested in this study are available in Supplementary Table S2.
  • Due to the multiplicative decrease of transfecting a two plasmid system vs a single GFP plasmid, transfection efficiency required further optimization. Seeding density, lipofectamine 3000 concentration, DNA concentration, light intensity, an additional wash step, and plasmid ratios were all varied to produce the optimal conditions for each cell ...
  • Sep 24, 2020 · Transgenic cells were made by transfecting 500 ng of plasmid via 1 μL of lipofectamine 3000 (L3000008; ThermoFisher) per well of 24 well plate containing 500 μL of Opti‐MEM (31985062; ThermoFisher) with 1X RevitaCell (A2644501; ThermoFisher).
  • Optimizing Transfection To obtain the highest transfection efficiency and low non-specific effects, optimize transfection conditions by varying cell density and Lipofectamine 2000 CD concentrations.

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If you have followed the given Lipofectamine 3000 protocol exactly as instructions are given, then average transfection efficiency should be 25-40%. However, few other factors that you can check...
Adherent 293T/17 cells were transfected in a 6-well plate with pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION ® (1:1 ratio, 2 μg/well, Mirus Bio) with the following reagents: TransIT ®-Lenti (3:1, vol:wt, Mirus Bio), Lipofectamine ® 2000 (3:1, Thermo Fisher Scientific), Lipofectamine ® 3000 ...

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Oct 05, 2015 · Formulation of Lipofectamine 2000 and Lipofectin. LIPOFECTIN is a 1:1 (w/w) mixture of N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA) and ...

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Feb 26, 2020 · The next day, f1 G8P PD plasmid (1.5 μg) were transfected into cells using Lipofectamine 3000 (Invitrogen). At 24 h post G8P PD transfection, plasmids encoding SaCas9 (0.75 μg) and EMX1 -targeting sgRNA or FANCY -targeting sgRNA (0.375 μg) were co-transfected into G8P PD -expressing HEK293T cells.