Lipofectamine™ 3000 Transfection Reagent

DNA transfection Mammalian cells - Primary cells Rat astrocytes

Experiment
DNA transfection Mammalian cells - Primary cells Rat astrocytes
Product
Lipofectamine™ 3000 Transfection Reagent from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

2.4. Transfection
Cells were seeded on either 24- or 96-well cell culture treated plates at a density to ensure ~80% confluent cultures at 24 hr after seeding. Typically 80,000 cells/cm2 surface area of the culture dish were seeded in cell-specific medium. Transfection using Lipofectamine 3000 was performed according to the manufacturer's protocol with a DNA to Lipofectamine ratio of 1 : 3 w/v. A transfection enhancer, the 3000 enhancer reagent (1 : 2, DNA : Reagent, w/v), was used along with the Lipofectamine 3000 transfection reagent for all transfections. Typically 100 ng and 500 ng of plasmid DNA were transferred to each well of the 96-well plate and 24-well plates, respectively. The standard complexation protocol was employed for Viromer RED according to the manufacturer's instructions. Briefly, Viromer RED was diluted (1 : 24 v/v) and the plasmid DNA was diluted independently to 18 ng/μL in the provided dilution buffer E. A 22 μL volume of diluted DNA was added to 4 μL of diluted Viromer RED and the mixture then was allowed to stay at room temperature for 15 min. For transfection, 100 ng of plasmid DNA, Viromer RED mixture, was added to each well of the 96-well plate by dispensing 6.7 μL per well. In specified experiments, 6 hr before transfection, the media was changed to either DMEM with antibiotics and without serum or DMEM with antibiotics and with serum. For quantitative evaluation of protease activity (as a measure of cell viability), luminometry, or fluorescence microscopy, cells were used 24 hr after transfection.

Full paper   Login or join for free to view the full paper.

Reviews

Lipofectamine™ 3000 Transfection Reagent from Thermo Fisher Scientific has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA transfection Mammalian cells - Primary cells Rat astrocytes using Lipofectamine™ 3000 Transfection Reagent from Thermo Fisher Scientific.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Lipofectamine™ 3000 Transfection Reagent below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA transfection Mammalian cells - Primary cells Rat astrocytes using Lipofectamine™ 3000 Transfection Reagent from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms