Description

PurMaFectin™ Transfection Reagent

Background

PurMaFectin™ Transfection Reagent contains highly advanced lipid nanoparticle technology. Also,  it provides superior transfection performance with improved application outcomes and reproducible results. Furthermore, PurMaFectin™ Transfection Reagent delivers superior transfection efficiency and improved cell viability. Do not add antibiotics to the medium during transfection as this causes cell death and interferes with transfection efficiency.

Special Features

• For the highest efficacy and the lowest dead cells, transfect cells at around 70~80% confluency. We strongly recommend starting the seeding by counting trypsinized cells so there will be high consistency among your experiments.
• Different cell types react differently to the number of passages when they are used for transfection. To minimize the impact of passage on efficiency of transfection, either use the cells with the same passage number. Alternatively,  use at least two different concentrations of transfection reagent as control in new transfect experiments before the condition is optimized.
• For HeLa cells, high level transfection requires the presence of serum in the medium during transfection. The amount of required serum depends on sequence and the length of plasmid. The exact level of serum should be determined empirically.
• We deplete endotoxin from our transfection reagent. The  endotoxin-contaminated DNA results in inefficient transfection and can cause high cellular toxicity. To deplete the endotoxin, we use PurMa^TM Endotoxin Elimination Column (Cat# P6H1216251). In this proprietary kit, Sepharose 6B covalently conjugates to an endotoxin-specific absorbent. The mentioned procedure will end with passing the reagent through 0.1µm filters 3X (included in the kit). The mentioned eliminates the residual endotoxin.
• Some serum-free medium might interfere with the transfection of cation ions and plasmid. We strongly recommend washing the cells with a conventional media such as DMEM prior to transfection. This will result in, eliminating serum can.  Afterwards, you can add serum to the media 2 hours post transfection.

User Manual

For complete protocol, click here:PurMaFectin_User Manual

References

1. Novel cationic liposomes for DNA-transfection with high efficiency and low toxicity. Paukku et al. Chem Phys Lipids. 1997 May 30;87(1):23-9. doi: 10.1016/s0009-3084(97)00020-0. PMID: 9219346.
2. A new efficient method for transfection of neonatal cardiomyocytes using histone H1 in combination with DOSPER liposomal transfection reagent. Kott et al. Cell Mol Genet. 1998 Jul;24(4):257-61. doi: 10.1023/b:scam.0000007128.56413.31. PMID: 10410680.