How do you transfect siRNA?
9 Tips for Optimal siRNA Transfection
- Use the most appropriate siRNA concentration.
- Prepare a suitable siRNA stock solution.
- Transfect healthy cells.
- Check serum quality.
- Know the target gene in and out.
- Always use positive and negative controls.
- Follow up the transfection reagent protocol.
What is the purpose of a transfection reagent?
Transfection Reagent Basics Transfection is the process of introducing nucleic acids into eukaryotic cells by nonviral methods. Using various chemical or physical methods, this gene transfer method enables the study of gene function and protein expression in a cellular environment.
Where are siRNA found?
siRNAs and their role in post-transcriptional gene silencing(PTGS) was discovered in plants by David Baulcombe’s group at the Sainsbury Laboratory in Norwich, England and reported in Science in 1999.
How long does it take to transfect Huh 7 cells?
The 24-well plate protocol to transfect HuH-7 cells is here: Plate 10,000-15,000 Huh-7 cells per well in 0.5 ml of complete growth medium 12-24 hours prior to transfection Wash with 1xPBS and add 0.5 ml of fresh growth medium Incubate transfection complexes at RT for 15-30 minutes
Which is an example of Huh 7 transfection?
HuH-7 is receptive to transfection with certain viruses. As an example, HuH-7 cells have been used to elucidate how HCV, the hepatitis C virus, interacts with hepatocytes in vivo.
What are the characteristics of Huh 7 cells?
Transfecting HuH-7 cells. HuH-7 cells are an immortalized hepatocellular carcinoma cell line originally resected from a male, 57 year old liver tumor. The HuH-7 cell line is an epithelial-like adherent cell line containing multiple mutations including a point mutation in p53. HuH-7 is receptive to transfection with certain viruses.
How are Huh-7 cells grown in a suspension?
HuH-7 cells grown in RWV suspension are incubated in a horizontally rotating cylindrical culture vessel that allows cells to grow in 3D aggregates, without the turbulence associated with conventional, stirred bioreactors. Cells cultured in RWV are grown much like cells of a fetus in a mother’s uterus.