Question

1. Which of the following answers is (are) true about size exclusion chromatography? a. large proteins...

1. Which of the following answers is (are) true about size exclusion chromatography?

a. large proteins elute first

b. small proteins interact more with the column

c. proteins larger than the cutoff size do not elute

d. will be used to purify GFP

2. Which of the following is (are) not true about ion exchange chromotagraphy?

a. positively charged proteins elute first

b. negatively charged proteins elute first

c. uncharged proteins elute first

d. hydrophobic proteins elute first

3. The use of an antibody to capture a specific target protein best describes ............ chromatography.

4. Which of the following is (are) true about hydrophobic interaction chromatograpgy?

a. will be used to purify GFP

b. low salt buffer elutes captured proteins

c. the column is hydrophilic and the buffer is hydrophobic

d. the column is uncharged

5. To achieve the goal of isolating..................... from the bacteria, the cells must first be lysed.

a.fluorescent protein

b. pGLO

c. plasmid

d.AraC protein

Lysis will be performed using...........................

a. lysozme

b. protease

c. catalase

d. hydrochloric acid

and...................

a.freezing-thawing

b. boiling

c. vortexing

d. sonicating

6. The bacteria that you previously transformed will be cultured overnight in nutrient broth. To grow only flourescent bacteria, the nutrient broth must provide?

a. arabinose

b. AraC

c. ampicilin

d. beta lactamase

e. pGLO

f. arabinose

7. If bacteria growing on the LB/amp plate were streaked on LB/amp/ara plates then?

a. they would fluoresce immediately

b. they would not flouresce because they do not have plasmid

c. they would die

d. they would not flouresce but their daughter cells would flouresce

e. they would flouresce once gene expression occured.

Homework Answers

Answer #1

1. Option C and D are the correct answer. Size exclusion chromatogrphy separation occurs on size basis and if the size of eluents will be more than the upper limit (cut-off size) then do not elute. It is also used to purify green fluoroscent protein by linking fluoroscent material covalently.

2. Option C and D are the correct answer. Ion exchange chromatogrphy deals with either anionic or cataionic materials.

3. Affinity chromatography.

4. Option A is correct. Here the column is hydrophobic which is initially treated with strongly ionic salt bugger.

5. Option C, A and B are correct options for respective parts of 5th question. Plasmid is the circular DNA and isolation of DNA always require lysis. Lysis can be performed by boiling with lysozyme.

6. Option E is the correct answer.

7. Option B is correct.

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