How do you take Lipofectamine?
How do you take Lipofectamine?
Mix Lipofectamine 2000 gently before use, then dilute the appropriate amount in 250 μl of Opti-MEM I Medium (or other medium without serum). Mix gently and incubate for 5 minutes at room temperature. After 5 minutes incubation, combine the diluted DNA with the diluted Lipofectamine 2000 (total volume is 500 μl).
How do you take Lipofectamine 2000?
Nucleic acid-Lipofectamine 2000 complexes can be added directly to cells in culture medium, in the presence or absence of serum. It is not necessary to remove complexes or change/add medium after transfection, but complexes may be removed after 4-6 hours.
What is the difference between lipofectamine 2000 and rnaiMaX?
Lipofectamine® rnaiMaX reagent is designed specifically for the delivery of sirna and mirna while Lipofectamine® 2000 reagent delivers Dna or sirna with excellent transfection performance for protein expression, gene silencing, and functional assays.
Can Lipofectamine 2000 be used siRNA?
invitrogen Lipofectamine 2000 Reagent is a proprietary formulation that facilitates highly efficient delivery of Invitrogen Stealth RNA molecules, short interfering RNA (siRNA) or plasmid DNA to mammalian cells for RNAi analysis (1, 2).
Which transfection reagent is best?
Under the tested conditions, ViaFect™ Reagent offered the best combination of transfection efficiency and low toxicity for most cell lines, making it an ideal choice when beginning transfection experiments with a new cell line.
What is the reagent protocol for Lipofectamine 2000?
Lipofectamine®2000 Reagent Protocol Outline A. Plate cells so they will be 70–90% confluent at the time of transfection. B. Prepare plasmid DNA-lipid complexes. C. Add DNA-lipid complexes to cells. Lipofectamine®2000 DNA Transfection Reagent Protocol See page 2 to view a typical DNA transfection procedure. Component 96-well 24-well 6-well
How to transfect NIH3T3 cells with Lipofectamine 2000?
Transfection of NIH3T3 cells, Hela, Swis 3T3, 293T with Lipofectamine 2000 Pre-warm 50ml of Optimum (stored in cold room at 4°C) ~10min. Add ~2µg of DNA to Eppendorf tube in the hood. Add 100µl of Optimum to the Eppendorf tube to dilute the DNA and mix by tapping.
How much Lipofectamine is used in a well?
DNA-lipid complex per well 10 μL 50 μL 250 μL Final DNA used per well 100 ng 500 ng 2500 ng Final Lipofectamine®2000 Reagent used per well 0.2–0.5 μL 1.0–2.5 μL 5.0–12.5 μL Visualize/analyze transfected cells Incubate cells for 1–3 days at 37°C. Then analyze transfected cells.
How long is Lipofectamine 2000 stable at room temperature?
After the 5 minute incubation, combine the diluted DNA with diluted Lipofectamine 2000 (total volume = 100 μl). Mix gently and incubate for 20 minutes at room temperature (solution may appear cloudy). Note: Complexes are stable for 6 hours at room temperature. Add the 100 μl of complexes to each well containing cells and medium.