5. (9 pts) Kyle is in the process of purifying a protein and would like to analy

Question

5. (9 pts) Kyle is in the process of purifying a protein and would like to analyze his first purification fraction (cell lysate) via SDS-PAGE to get a sense of how many proteins are present in this fraction. He wants to run four different concentrations of this fraction (like we did in the SDS-PAGE experiment). Kyle plans to prepare three dilutions with concentrations of: 1.2 ug/uL, 0.6 ug/uL, and 0.12 ug/uL. He has decided that he'll make volume of these dilutions 100 uL. Describe how Kyle should prepare these samples using serial dilutions. He has reserved 100 uL of this first purification fraction (concentration- 3.2 ug/uL). (6 pts) stuc have hrst, 3.2 Assume that Kyle loads 15 uL of each of his dilutions onto the gel, how many micrograms of protein will he be loading? (3 pts) Dilution number Total ug of protein loaded into gel 2

Explanation / Answer

A -

Inintial concentration Volume added Final Concentration Volume of Dileunt Total volume Dilution Factor 3.2 µg/µl 75 µl 1.2 µg/µl 125 200 µl 2.66 times 1.2 µg/µl 100 µl 0.6 µg/µl 100 200 µl 2 times 0.6 µg/µl 40 µl 0.12 µg/µl 160 200 µl 5 times

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