Midterm Review for Monday Bio Molec Lab 140A. - Midterm...

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Midterm Review for Monday 1. Molecular was to determine the bands. 2. Uninduced from most proteins, proteins didn’t get expressed 3. Induced expression of DHFR (an enzyme), dna inserted into plasmid (producing protein isolation) 4. Insoluble/Soluble is dividing the proteins in half. Some proteins of interests, they are all clumped together with no space between them, water can’t get into them. 5. Flow through → we mixed the sample with resin, and histidine were binding to the small molecules. All soluble proteins except for 1 6. Eluate → adding metizole, eluation buffer had high solution of ____
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Unformatted text preview: flowing through the sample 7. Desalted eluate ---> lack of metizole, flowing through the metizole 8. All of the wells mixed with Lamelli buffer to ro run together, the charge will not dictate how fast they can run through, only by size. ALL RAN AT THE SAME TIME, separated by size not charge. 9. How did we isolate dna protein? By “GSt -dhfr-HIS”. Know the functions of DHFR. “GST” HELPED TO BE SOLUBLE added GST to be expressed. 10.Histidine helps bind to the column. 11.All proteins in e coli will bind more than histidine....
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