how to calculate loading percentage

Calculating loading percentage is an important aspect of any experimental design. Whether doing Western blotting or PCR, it's important to know how much of your sample was loaded onto the gel or plate. Loading percentage refers to the amount of protein or DNA loaded relative to the total volume of the sample. In this article, we will explore how to calculate loading percentage for different experimental techniques.

Western Blotting

For Western blotting, loading percentage is calculated based on the amount of protein loaded onto the gel. The loading percentage is the weight of the protein sample divided by the total weight of the sample loaded onto the gel. For example, if you loaded 50 micrograms of protein in 100 micrograms of the total sample, then the loading percentage would be 50%.

SDS-PAGE Gel Electrophoresis

In SDS-PAGE gel electrophoresis, loading percentage refers to the amount of protein loaded relative to the total volume of the sample. This can be calculated using the formula:Loading Percentage = (Volume of Sample Loaded ÷ Total Volume of Sample) x 100For example, if you loaded 20 microliters of a 100 microliter sample, then the loading percentage would be 20%.

PCR

In PCR, loading percentage refers to the amount of DNA loaded relative to the total volume of the sample. This is calculated using the formula:Loading Percentage = (Amount of DNA Loaded ÷ Total Volume of Sample) x 100For example, if you loaded 100 nanograms of DNA in 200 microliters of total sample, then the loading percentage would be 0.05%.

Flow Cytometry

In flow cytometry, loading percentage refers to the amount of cells loaded relative to the total volume of the sample. This can be calculated using the same formula used for PCR:Loading Percentage = (Number of Cells Loaded ÷ Total Volume of Sample) x 100For example, if you loaded 1 million cells in a total sample volume of 500 microliters, then the loading percentage would be 0.2%.

Conclusion

Calculating loading percentage is an important step in many experiments. It is important to know the loading percentage so that you can accurately interpret your results. By using the formulas outlined in this article, you should be able to easily calculate the loading percentage for a variety of experimental techniques. Remember, accuracy is key, so make sure to measure your samples carefully.