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For example, if you plate 1x 10 7 cells and count 1000 colonies, the transformation efficiency is: (1000/1x 10 7) x 100 = 0.1% Alternatively, CFUs can be reported per microgram of DNA used for the transformation. This can be calculated by multiplying the number of colonies by the volume of the culture plated and dividing by the amount of DNA used.
Three plates are needed for each dilution series, for statistical reasons an average of at least 3 counts are needed. The surface of the plates need to be sufficiently dry to allow a 20μl drop to be absorbed in 15–20 minutes. Plates are divided into equal sectors (it is possible to use up to 8 per plate).
The "dilution factor" is an expression which describes the ratio of the aliquot volume to the final volume. Dilution factor is a notation often used in commercial assays. For example, in solution with a 1/5 dilution factor (which may be abbreviated as x5 dilution ), entails combining 1 unit volume of solute (the material to be diluted) with ...
A dilution of the cells to be counted is prepared and mixed with Trypan blue, this is normally the stain of choice because it is taken up by dead cells and actively excluded from live cells. Once the cells have been stained, they are counted using a hemocytometer, then a calculation is carried out to the original concentration of live cells. [1]
Plating Efficiency is the number of cells that grow into colonies per 100 cells inoculated. That is, it is the proportion of cells that attach and grow to the number of cells originally plated, expressed as a percentage. PE can be determined by the following formulae:
These cell types can be subcultured by simply taking a small volume of the parent culture and diluting it in fresh growth medium. Cell density in these cultures is normally measured in cells per milliliter for large eukaryotic cells, or as optical density for 600nm light for smaller cells like bacteria. The cells will often have a preferred ...
One of the most important features of chemostats is that microorganisms can be grown in a physiological steady state under constant environmental conditions. In this steady state, growth occurs at a constant specific growth rate and all culture parameters remain constant (culture volume, dissolved oxygen concentration, nutrient and product concentrations, pH, cell density, etc.).
A serial dilution is the step-wise dilution of a substance in solution, either by using a constant dilution factor, or by using a variable factor between dilutions. If the dilution factor at each step is constant, this results in a geometric progression of the concentration in a logarithmic fashion.