What is lipofectamine transfection?

Lipofectamine or Lipofectamine 2000 is a common transfection reagent, produced and sold by Invitrogen, used in molecular and cellular biology. It is used to increase the transfection efficiency of RNA (including mRNA and siRNA) or plasmid DNA into in vitro cell cultures by lipofection.

What is Lipofectamine Plus reagent?

Lipofectamine LTX Reagent is a plasmid transfection reagent that offers a balance of potency and gentleness for your cells, resulting in high transfection efficiencies and viabilities.

How do you mix Lipofectamine?

Mix Lipofectamine 2000 gently before use, then dilute 1 μl in 50 μl Opti- MEM I Reduced Serum Medium. Mix gently and incubate for 5 minutes at room temperature. Note: Proceed to Step c within 25 minutes. c.

What is LTX transfection?

Lipofectamine™ LTX Reagent is a plasmid transfection reagent that offers the right balance of potency and gentleness for your cells, resulting in the highest possible transfection efficiencies and viabilities.

Is lipofectamine toxic to cells?

2.3. Although Lipofectin showed the lowest toxicity to HepG2 cells (89.54% viability), it displayed the lowest transfection efficacy too (8.29%). Generally, HepG2 cells displayed resistance to the toxicity of majority of the reagents tested, with Lipofectamine 3000 (70.59 % viability), being the most toxic reagent.

What is plus reagent?

PLUS™ Reagent is a proprietary reagent for pre-complexing DNA that enhances cationic lipid-mediated transfection of DNA into many cultured eukaryotic cells.

How do you add lipofectamine to a cell?

  1. Label two tubes and add 125 ul of Opti-MEM to each.
  2. In the first tube add ~ 5 ul of L3000 – vortex.
  3. In the second tube add ~ 2.5 ug of DNA and then add ~ 5 ul P3000 and mix by pipetting (do not vortex)
  4. Add the contents of the second tube into the first tube.
  5. Incubate for 10 – 15 minutes at room temp.
  6. Add to cells.

When should I change my medium after transfection?

A complete media change can be performed 5 – 24 hours after transfection for very sensitive cells. For most cells, we recommend the media be changed only at 48 hours post-transfection until protocol optimization requires this extra media change. A complete media change should be performed at 48 hours post-transfection.

What are transfected cells?

Transfection is a procedure that introduces foreign nucleic acids into cells to produce genetically modified cells. Transfection is a powerful analytical tool for study of gene function and regulation and protein function.

How do you increase transfection efficiency of Lipofectamine 3000?

  1. The day before transfection, Plate 4×105 cells per well in 0.5 mL of complete growth medium.
  2. For each well of cells to be transfected, dilute 1 μg of DNA in 50 μL of Opti-MEM , and 2.5 μL of P3000 in 50 μL of Opti-MEM in separated tube.
  3. Mix gently and incubate 20 minutes at room temperature.