Purification technique essay

i have completed the calculation for this lab but i don;t have time to finish the writing part. the writing part is to answer 3 questions depending on the data we get from the calculation. I will upload my data exell sheet and assignment requirement. 1. Which purification technique lost the most LDH? Discuss this in the context of the LDH recovery for each purification step. You should calculate the percent recovery of LDH (the fraction of the LDH that went into that step that was recovered) for each step and provide it here. Which steps recovered the LDH better and which steps lost a lot? This should be at least 100 words but not more than 200 words. 2. Which technique gave the highest degree of purification of the LDH? Tell how you are assessing purification. To which columns are you referring to determine the amount of purification achieved in each step? Specific activity is most important but what else do you need to look at? You should discuss purification in terms of the fold-purification provided by each step. Fold-purification is the specific activity achieved by a purification step divided by the specific activity of the previous step. Be sure to answer the question but place it in the context of the purification achieved by each technique (if you simply list numbers without placing them in context you will lose points). This should be 150 – 300 words. 3. How would you modify this purification strategy to maximize the yield and purity of LDH? Which steps in the original purification strategy worked well and which did not? What would you change to get better recovery and purification of LDH? Give each step/technique of your modified strategy and describe what is accomplished by that step. You can change the parameters or way that any individual technique is done. You can also change the of the techniques used remove a technique and even add a new technique that you have Purification Intermediate Relative Activity (U/ml) Total LDH Activity Units (U) Protein Concentration (mg/ml) Total Protein (mg) Specific Activity (U/mg) Clarified Homogenate 65% Cut Pellet Affinity-Purifie d LDH LDH Final Product not performed. Just be sure that you can provide a rationale for each step in your new purification strategy based on the purification table data. How will that modification improve the purification? (Be sure you understand what is meant by this; ask if you don’t!) You should provide numbers from the purification table in making your arguments here. Keep in mind that the initial crude homogenate is not listed in the purification table and the clarified homogenate is given as the starting material. This is because the crude homogenate is very difficult to work with; its turbidity interferes with the assays and the LDH is rapidly degraded by proteases. Even though you didn’t measure it you may assume the initial centrifugation you performed to isolate the cellular fraction containing LDH achieved a good degree of purification from the crude homogenate. You should account for this in your new purification strategy. This should be one to two pages. Put some careful thought and consideration into answering these questions. Your score on this is based on the quality of your writing including how well you make your arguments and provide a careful rationale for your answers.

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