a: ands b: alpha-MF/malE : bla PvuII 10153 bp Hpal PvulI Pull 15 points. The pla
ID: 3164890 • Letter: A
Question
a: ands b: alpha-MF/malE : bla PvuII 10153 bp Hpal PvulI Pull 15 points. The plasmid shown above was digested (cut) with the restriction enzyme, Hpa !. How many bands will be seen on an agarose gel as a result of this digestion? The same molecule was cut with Pvull. How many bands will be seen in this case? How many bands will be seen if the molecule is digested with the two enzymes together? Please indicate on the molecule above the fragment that will be the LARGEST after this double digestion Please indicate on the molecule shown the band that will run the furthest (fastest) during agarose gel electrophoresis. (You can circle it if you want). 1. (You can circle it, if that is easier) 15 points. Our E. coli transformation experiment with plasmid pRS315 worked very nicely! All of us got colonies on our ampicillin plates, just as we hoped. But sometimes, experiments can go wrong. For each of the following mistakes, please state what the plates would have looked like in the end. Explain VERY briefly. 2. a. We forgot to add the plasmid DNA to the cells. b. The person who made the ampicillin plates forgot to put in the ampicillin. c. The person who made the ampicillin plates put in the wrong antibiotic (e.g., tetracycline instead of ampicillin.)Explanation / Answer
Answer 1: The plasmid when cut with HpaI wiill show three bands on agarose gel
The plasmid when cut with PvuII wiill show three bands on agarose gel
The plasmid when cut with both HpaI and PvuII wiill show six bands on agarose gel
Largest band will be the one having HpaI restriction sites on both end and having an arrow containing b sequence.
Smallest fragment will run the fastest and it is the one having at one end HpaI and PvuII after a sequence
Answer2: a) if plasmid DNA is not added in the cells, no colonies will be seen since DNA is needed for transformation to occur.
b) If ampicillin is not added, then plates will be full of colonies both that are transformed and untransformed
c) If tertracycline is added instead of ampicillin no colonies will be seen since plasmid is resistant to ampicllin not tetracycline and all colonies will die.
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