What is the difference between lipofectamine 2000 and RNAiMAX?

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.

What is Lipofectamine made of?

Lipofectamine consists of a 3:1 mixture of DOSPA (2,3‐dioleoyloxy‐N‐ [2(sperminecarboxamido)ethyl]‐N,N‐dimethyl‐1‐propaniminium trifluoroacetate) and DOPE, which complexes with negatively charged nucleic acid molecules to allow them to overcome the electrostatic repulsion of the cell membrane.

What is forward and reverse transfection?

The key difference between forward and reverse transfections is the cell density at the time of plating. Cells lose a day of doubling in a typical reverse transfection protocol; therefore, approximately twice the number of cells are recommended compared to a forward transfection.

How do you transfect cells with siRNA?

9 Tips for Optimal siRNA Transfection

1. Use the most appropriate siRNA concentration.
2. Prepare a suitable siRNA stock solution.
3. Transfect healthy cells.
4. Check serum quality.
5. Know the target gene in and out.
6. Always use positive and negative controls.
7. Follow up the transfection reagent protocol.

What is the difference between lipofectamine 2000 and 3000?

Lipofectamine 3000 reagent yields higher transfection efficiencies than Lipofectamine 2000 reagent when tested in a variety of cell lines.

How does siRNA affect gene expression?

The siRNA-induced post transcriptional gene silencing starts with the assembly of the RNA-induced silencing complex (RISC). The complex silences certain gene expression by cleaving the mRNA molecules coding the target genes. This cleavage results in mRNA fragments that are further degraded by cellular exonucleases.

Is Lipofectamine toxic to cells?

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 Lipofectamine RNAiMAX?

Lipofectamine® RNAiMAX is a proprietary RNAi-specific cationic lipid formulation designed specifically for the delivery of siRNA and miRNA into all cell types.

Is reverse transfection better?

Advantages and disadvantages. The advantages of reverse transfection (over conventional transfection) are: The addition and attachment of target cells to the DNA-loaded surface can lead to a higher probability of cell-DNA contact, potentially leading to higher transfection efficiency.

What are transfected cells?

Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. Transfection of animal cells typically involves opening transient pores or “holes” in the cell membrane to allow the uptake of material.

How do you dissolve siRNA?

General considerations: Dissolve siRNA in sterile water and prepare 5 µl aliquots, store at –80°C. For use, dilute siRNA in siRNA dilution buffer (diluted siRNA is not stable, use only once). 5x siRNA buffer aliquots (Dharmacon) are stored at 4°C in the cell culture lab.

Can I Vortex siRNA?

Incubate the diluted siRNA mixture at room temperature for 5 min. Important: do NOT vortex the diluted siRNA mixture. Incubate at room temperature for 5-30 minutes (5 minutes is typically sufficient).

How is Lipofectamine rnaimax used in cell therapy?

Lipofectamine RNAiMAX is proprietary animal origin-free, chemically defined RNAi-specific cationic lipid formulation designed specifically for delivery of siRNA and miRNA into all cell types. Superior transfection efficiency requiring lower RNAi concentrations, leading to more effective gene knockdown with minimal nonspecific effects

Which is the best Lipofectamine for reverse transfection?

Invitrogen Lipofectamine RNAiMAX is a proprietary formulation specifically developed for the transfection of siRNA and Invitrogen Stealth RNAi duplexes into eukaryotic cells. Lipofectamine RNAiMAX provides the following advantages: Reverse transfection and forward transfection protocols can be used for most cell lines tested.

Which is the best concentration for rnaimax reverse transfection?

To obtain the highest transfection efficiency and low non-specific effects, optimize transfection conditions by varying RNAi duplex and Lipofectamine RNAiMAX concentrations. Test 0.6-30 pmol RNAi duplex (final concentration 1-50 nM) and 0.5-1.5 µl Lipofectamine RNAiMAX for 24-well format.

Which is the best rnaimax reagent for Knockdown experiments?

With its ability to transfect a broad range of cell types, Lipofectamine RNAiMAX reagent, combined with the Invitrogen BLOCK-iT Alexa Fluor Red Fluorescent Control, gives you the most versatile approach to all of your knockdown experiments (Table 1, Figure 3).