Biology > Lab Report > Multi-column purification of rGFP University of Texas, Dallas - BIOL 33803380 Lab 8 online F20 lab r (All)

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Multi-column purification of rGFP University of Texas, Dallas - BIOL 33803380 Lab 8 online F20 lab report

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Experiment 8 – Lab Report Multi column Purification of rGFP To save time, simply edit this word document by inserting your answers after each question/space. Please denote your answers by using b ... old, italics, or colored font. Do NOT renumber/reorganize the questions. “Showing your work/calculations” can be achieved by typing it out, pasting in a clear image of your handwritten work, or using handwritten annotation on the document (if you have the technology to do so). Insert graphs (handwritten on graph paper or electronically constructed with horizontal/vertical gridlines) as a clear image with appropriate titles, labeled/scaled axes, and standard deviation bars when appropriate. 1. My assigned data set for experiment 4 was: (1pt) Which two fractions did you pool from column 1 (Ni+2 agarose) to apply to column 2 (anion exchanger)? E3 and E4 were pooled and applied together to column 2 anion exchanger because these fractions had the highest RFUs. 2. My assigned data set for experiment 4 was: (3pts) What was the amount of rGFP activity that you loaded onto column 2? Show your calculations for credit. Fraction E3 pool 1 RFUs: 6,021 RFU/200ul = X RFU/ 400ul loaded X RFU = 12,042 RFU Fraction E4 pool 1 RFUs: 4,645 RFU/200ul = X RFU/ 400ul loaded X RFU = 9,290 RFU Pooled RFU = (12,042+9,290)/2 = 10,666 RFU 3. (3pts) Based upon the protocol, if the highest fluorescence was found in fraction F32, does rGFP carry a net positive, net zero, or net negative charge? Explain your reasoning for full credit. rGFP carries a net negative charge because fraction 32 is the result of the uses of an excess of NaCl salt, Cl- specifically, to outcompete the negative net charged rGFP binding to the positive beads of the anion exchange column [Show More]

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