Question

10mg/1ml pure herring sperm diluted with 10ml sterile distilled water to C1 of 1000µg/ml. C1 Concentration of stock (1000µg/ml) V1 Volume of stock needed (µl) C2 Final Concentration (µg/ml) O 100 80 75 70 60 50 30 25 5 V2 Final volume (2,000μl) 2,000 2,000 2,000 2,000 2,000 2,000 2,000 2,000 2,000 1,000 1,000 1,000 1,000 1,000 1,000 1,000 1,000 1,000 Volume of H₂0 required (μl) Use water as a diluent and blank Mix each standard by inversion and measure the absorbance at 260nm (A260) with a UV - compatible cuvette. Graph your standard curve. On the graph include coefficient of regression (R²) and line graph equation (Y = mx + C).​

Answer 1

To create a standard curve using the provided data, you will need to dilute the pure herring sperm stock solution to various concentrations. The table below shows the calculations for each dilution:

C1 Concentration of Stock (µg/ml)V1 Volume of Stock Needed (µl)C2 Final Concentration (µg/ml)V2 Final Volume (µl)Volume of H₂O Required (µl)100010010002000190010008080020001920100075750200019251000707002000193010006060020001940100050500200019501000303002000197010002525020001975100055020001995

To prepare each standard, you will take the corresponding volume of the stock solution and dilute it with the calculated volume of sterile distilled water. For example, to prepare the first standard with a final concentration of 1000 µg/ml, take 100 µl of the pure herring sperm stock solution and add 1900 µl of sterile distilled water.

Once you have prepared each standard, you can measure the absorbance at 260nm (A260) using a UV-compatible cuvette. Record the absorbance values for each standard.

To create the standard curve, plot the concentrations (C2) on the x-axis and the absorbance values (A260) on the y-axis. Fit a linear regression line to the data points. The equation of the line will be in the form Y = mx + C, where Y is the absorbance, m is the slope of the line, x is the concentration, and C is the y-intercept.

Calculate the coefficient of regression (R²) to assess the linearity of the curve. The R² value ranges from 0 to 1, with 1 indicating a perfect fit.

Please note that I cannot make graph for you directly in this text-based format. You will need to input the data into a graphing software or spreadsheet program to create the graph, calculate the regression line equation, and determine the coefficient of regression (R²).