This repository can be found at https://github.com/davidalliband/protein_dilution_calculator
This tool uses Python to quickly calculate the volumes to be combined from various total protein solutions (such as a BCA assay) to create a solution containing a constant protein quantity with buffer and loading dye.
To use this tool you must format your CSV file correctly. Column A should contain your samples (e.g. Sample 1, Sample 2, Sample 3, etc.) and be entitled "Content". Column B should contain the total protein of the sample in ug and be entitled "Total_protein". This will be used to calculate the volume of each Sample required to create a working solution containing your desired quantity of protein. An example CSV file is included with this tool (BCA_example.csv) with six samples with varying quantities of total protein in mg per ml.
To use this tool:
- Download all the contents of this repository including the script and example data into a single directory.
- Open the terminal and change directory to the folder in which the script is located.
- Enter "python western_dilution_calculator.py" in the terminal and hit return.
The script will then prompt you to enter certain information:
- The filepath of your data CSV. If the file is located in the same directory then simply enter the filename. For example, using the example data you would enter "BCA_example.csv".
- The volume of your total protein value in ul. For example, if your total protein is in mg per ml then enter 1000.
- Dilution factor of your working solution. For example, if calculating from mg per ml then 20000 would give 20ug of protein and 15000 would give 15ug of protein.
- Total solution volume in ul per replicate, for example, 35.
- Desire loading dye fraction. For example, for a 1 in 5 dilution enter "5".
- How many replicates you wish to create a working solution for.
Finally, you will be asked to enter the a filename for your new CSV file containing all the new data which will be exported to the same folder as the script.
This script performs a number of calculations in order to give the required volumes for the working solution. These are as follows:
- Sample volume to pipette per replicate to give the desired protein quantity = Dilution factor / Total protein
- Loading dye volume to pipette per replicate = Total solution volume per replicate / Loading dye fraction
- Buffer volume to pipette per replicate = Total solution volume per replicate / (Loading dye volume per replicate + Sample volume per replicate)
- Total solution volume = Number of replicates x Total volume per replicate
- Total sample volume to pipette = Number of replicates x Sample volume per replicate
- Total buffer volume to pipette = Number of replicates x Buffer volume per replicate
- Total loading dye volume to pipette = Number of replicates x Loading dye volume per replicate