How do you transfect HeLa cells?
Transfection of HeLa S3 Cells Plate 4 x 104 cells per well in 0.5 ml of complete growth medium. Cell density should be 50~80% confluent on the day of transfection. For each well of cells to be transfected, dilute 0.25 μg of DNA into 100 μl of Opti-MEM® I Reduced Serum Medium without serum.
How do you take lipofectamine?
Add the oligomer-Lipofectamine 2000 complexes to each well containing cells and medium. Mix gently by rocking the plate back and forth. Incubate the cells at 37°C in a CO2 incubator for 24-96 hours until you are ready to assay for gene knockdown. Medium may be changed after 4-6 hours.
Is lipofectamine stable transfection?
Lipofectamine® 3000 reagent leverages our most advanced lipid nanoparticle technology to enable superior transfection performance and reproducible results. It delivers exceptional transfection efficiency into the widest range of difficult-to-transfect and common cell types with improved cell viability.
Does lipofectamine expire?
According to ThermoFisher, lipofectamine 3000 can be used until 2 years after the manufacturing date if stored properly (stored in a 4 degree fridge should be fine). The Optimem is sensitive to expiration. I would recommend using the new reagents for your experiment. Hope this helps!
Is HeLa easy to transfect?
Even though HeLa cells are very hardy, the HeLa cell line is fairly easy to transfect and very receptive to viral vectors, making it a popular research tool in virology.
Are HeLa cells hard to transfect?
HeLa, one of biology’s workhorse cell lines, is refractory to high transfection efficiency using chemical methods. Cell lines that have been cultured too long or transfected multiple times might resist transfection as well.
What is the protocol for Lipofectamine 2000?
Lipofectamine®2000 DNA Transfection Reagent Protocol Transfect cells according to the following chart. Volumes are given on a per-well basis. Each reaction mix is sufficient for triplicate (96-well), duplicate (24-well), and single well (6-well) transfections, and accounts for pipetting variations.
How much Lipofectamine do you put in a 24-well transfection?
Test 10-50 pmol RNA and 0.5-1.5 μl Lipofectamine 2000 for 24- well format. Depending on the nature of the target gene, transfecting cells at higher densities may also be considered when optimizing conditions. Use the following procedure to transfect DNA into mammalian cells in a 24-well format.
How much DNA is in Lipofectamine 2000 reagent?
Component 96-well 24-well 6-well Final DNA per well 100 ng 500 ng 2500 ng Final Lipofectamine®2000 Reagent per well 0.2–0.5 μL 1.0–2.5 μL 5.0–12.5 μL Co-Transfection of Plasmid DNA and siRNA
Is HeLa transfection safe to use?
HeLa transfection is usually pretty standard in terms of protocols – just make sure your nucleic acid concentration is conforming to that of the protocol you’re using and you should be fine.