I am unsure how to show the calculations for the following. Please help. PROCEDU

Question

I am unsure how to show the calculations for the following.

Please help.

PROCEDURE 1. Switch on the spectrophotometer to give it time to warm up before you use it. It wi take about 20 minutes for it to warm up. 2. You will first prepare a series of protein "standards" for generating a standard curve. You'll use the standard curve with the spectrophotometer readings for your samples to estimate the levels of protein in your samples. To make the standards, you'll be given a vial containing bovine serum albumin (BSA) protein at a concentration of 1 mg/ml. Using this stock solution and sterile H20 (not tap water!) as a diluent, prepare tubes containing the following concentrations of BSA: (200 ul total volume each): Standard 1: 125 ug/ml BSA Standard 2: 62.5 ug/ml BSA Standard 3: 31.25 ug/ml BSA Standard 4: 15.625 ug/ml BSA CALCULATIONS: Show your calculations for making these samples to your TA before you start.

Explanation / Answer

Answer:
Based on the given information:

The stock concentration of the BSA solution is: 1mg/ml or 1000ug/ml (*** conversion factor 1mg = 1000ug).

After each dilution, vortex the sample to ensure complete mixing.

Sample Conc. Required ug/ml Fold dilution (x) Volume of Sample added (ul) Volume of diluent (sterile water) added ul) Final Conc. ug/ml Final volume (ul) 1000 (stock) 1000 Dilution1 125 =1000/125 = 8x 50 ul from stock 350ul 125 400 200ul used for serial dilution, 200ul left Serial Dilution 1 62.5 =125/62.5 = 2x 200ul from Dilution1 200ul 62.5 400 200ul used for serial dilution, 200ul left Serial Dilution 2 31.25 =62.5/31.25 = 2x 200 ul from Serial Dilution1 200ul 31.25 400 200ul used for serial dilution, 200ul left Serial Dilution 3 15.625 =31.25/15.625 = 2x 200ul from Serial Dilution2 200ul 15.625 400 400ul left

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