Table of Contents
Can I transfect cells twice?
All Answers (7) Yes, it can be transfected, in principle.
How do you show transfection efficiency?
Spectrophotometry A spectrophotometer is an instrument that measures light intensity as a function of color or wavelength. ß-galactosidase assays can be used to determine transfection efficiency by measuring colorimetric changes associated with ß-galactosidase activity.
How does plasmid size affect transfection efficiency?
5) were dependent on the size of the plasmid DNA, either supercoiled or linearized. Therefore, the topology of the transfected DNA molecules determines the level of gene expression but small DNA molecules have higher transfection efficiencies than larger ones.
How do you count cells for transfection?
Transfection efficiency can be calculated by determining the number of cells that exhibit transfected DNA over the total number of cells in the sample. The number of cells can be calculated under the microscope or by flow cytometry using a reporter system.
How confluent should cells be for transfection?
Confluency. As a general guideline, transfect cells at 40–80% confluency. Too few cells will cause the culture to grow poorly without cell-to-cell contact. Too many cells results in contact inhibition, making cells resistant to the uptake of foreign DNA.
Why is 37 °C the optimal temperature for transfection?
This phenomenon could be because 37 °C is the optimal culture temperature for mammalian cells ( Wang et al., 2017a ). Meanwhile, chemical transfection appeared to be less appealing than viral and physical transfection for transfecting primary cells, especially in human primary stem cells.
Can flow cytometry improve transfection efficiency in cells?
A novel rapid and reproducible flow cytometric method for optimization of transfection efficiency in cells. PLOS ONE. 2017;12(9):e0182941. doi: 10.1371/journal.pone.0182941.
What is transfection and how does it work?
Transfection is a process by which foreign nucleic acids are delivered into a eukaryotic cell to modify the host cell’s genetic makeup ( Kim & Eberwine, 2010; Chow et al., 2016 ).
Does serum starvation improve transient transfection efficiency in embryonic stem cells?
Wallenstein et al. (2010) Wallenstein EJ, Jeffrey B, Schloss RS, Yarmush ML. Serum starvation improves transient transfection efficiency in differentiating embryonic stem cells. Biotechnology Progress. 2010;26(6):1714–1723. doi: 10.1002/btpr.472.