Samantha wants to purify \"protein X\" from e. coli cells. She extracts proteins

Question

Samantha wants to purify "protein X" from e. coli cells. She extracts proteins from e. coli, runs them through a column that selectively binds only protein X, and then extracts the remaining protein X. She wants to double check her work, so she loads the following into a polyacrylamide ge Lane 1: protein standards (MW) Lane 2: The protein extract (what was loaded into the column) Lane 3: Proteins that come out with the wash buffer (proteins that did not bind to the (75 kD) ok column) Lane 4: Elute (the solution with purified protein, which comes out at the end) 20 CKOA) Briefly answer the following questions about the protein gel on the right: 6) What was the purpose of lane 1 (protein standards)? do th rest lan Be enurlto this me 7) Do you think she purified a protein? Was there mostly one type of protein in the elute" sample?]? What was the approximate size of the protein? ?? oni 8) What kind of stain did she use to visualize all the dijferent pr stain for full credit)? oteins in the gel (give either the common or chemical name of the that bind to the beginning 9) f Samantha wants to look at the gene that codes for protein X, she should design DNL shael and endof the gene, into and copy the X gene using a PCR reaction, Then she should see if her PCR reaction works by loading her sample gel where the DNA is stained with T -

Explanation / Answer

8. Coomassie blue R 250 dye is used to visualize the different proteins in the gel.

9. If Samantha wants to look at the gene that codes for protein X, she should design Primers that bind to the beginning and end of the gene, and copy the X gene using a PCR reaction. Then she should see if her PCR reaction works by loading her sample into polyacrylamide gel where the DNA is stained with Ethidium bromide.

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