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You are asked to run an SDS-PAGE analysis of samples from an affinity chromatography experiment
performed by your TA. A recombinant protein with a hexahistidine tag was captured in a nickel- agarose bead matrix and eluted off the column with 75mM imidazole. Your TA was hoping to make about 2 mg/mL of the protein. You are given the following samples to prepare and load into an SDS-PAGE gel :
· 10 uL 75 mM Imidazole
· 10 uL pure recombinant protein at 2 mg/mL
· 10 uL recombinant protein in 75 mM imidazole (elution fraction)
1. Describe how you would prepare the three samples to load into the gel.
2. Label the gel lanes with the appropriate contents. The recombinant protein should exhibit one band at 66,000 Daltons
3. Compare the elution lane to the positive control lane. How do you know the recombinant protein present in the elution sample? How pure is the elution sample compared to the control?
4. What assumption can you make about the amount of recombinant protein in the elution sample? What assay can you perform to find the accurate concentration of the elution sample?
Suppose that the each replication fork moves at a rate of 500 base pairs per second, how long would it take to replicate the E. coli chromosome from a single origin of replication?
What changes took place from one group to another. What advantages did each progressive group have over the last? Will you categorize each as more sucessful than the last.
Determine the genotypes of parental, recombinant and if applicable double and triple recombinant progeny from the testcrosses of following individuals.
Discuss how the different amino acids contribute to protein structure. You do not need to discuss the role of amino acids in metabolism in this question.
Complete each of statements given below to identify whether the solution inside each bag washypertonic, hypotonic or isotonic.
Understand and discuss how the flow of nutrients differs from the dlow of energy in an ecosystem.
Your favorite patient has just had surgery. After surgery, Bactroban was applied. After a few days, you notice an infection around incision. Even though more bactroban is applied, the infection persists.
Suppose when performing 2-proteins, one having a molecular weight of 10000 and the other having molecular weight of 100000. both are globular and of similar spherical shape.
As a Nursing student in Advanced Human Anatomy, you were able to observe a total hip replacement. While the O.R. personnel where prepping the patient for the surgical procedure.
Tay-Sachs is a recessive lethal disease in which there is neurological deterioration early in life. The disease is rare in the population overall but is found at relatively high frequency in Ashkenazi Jews from Central Europe.
Within the two groups defined by bone mass, smaller skeletons have bones with evidence of epiphyseal plates, but the larger skeletons have only a thin line where the epiphyseal plates should be. Give the age group and gender of the four individuals i..
A body was discovered and diagnosed as having died due to oxygen deprivation. Explain why the toxicology tests included a test for cyanide poisoning.
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