Answer-Key---Recombi_36751

Answer-Key---Recombi_36751 - BIOTECHNOLOGY PROBLEM 1 Using...

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BIOTECHNOLOGY PROBLEM 1 Using the following mammalian DNA sequence and the restriction enzyme, Taq I, find the restriction sites and indicate where the cuts occur. Taq I : 5’ T*CGA 3’ 3’ A GC*T 5’ C T * C G A T T A T * C G A A C C T T C A T * C G A T T * C G A G C C T T C G G A G C * T A A T A G C * T T G G A A G T A G C * T A A G C * T C G G A A G C Circle the fragments generated by the restricition enzyme and label A, B, C, ….and so on. How many total fragments are generated? 5 fragments: (A-6 nucleotides; B-14; C-22; D-10; E-18) If these fragments were separated using gel electrophoresis, indicate below using the letter designation previously assigned, to depict the pattern: Towards which direction do the DNA fragments migrate ? Why ? Positive pole; DNA is negatively charged (the phosphate groups in DNA) Part 2. One of the fragments above contains a gene of interest you wish to isolate. You have a test tube containing the DNA fragments formed from the restriction enzyme reaction in Part 1. In order to execute the next phase of study, you are then provided a suspension containing bacterial plasmid (shown below), as well as a tube of DNA ligase. You are going to form recombinant DNA molecules. How do you do this? What is DNA ligase for ? - + A B C D E C E B D A Cut the plasmid with restriction enzyme; the same enzyme that you cut the mammalian DNA above, i.e., Taq I. Why? This generates the same compatible ends so that the cut plasmids and mammalian DNA fragment ends can complimentary base pair to form
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This note was uploaded on 04/15/2010 for the course BIOL 1362 taught by Professor Loeblich during the Spring '08 term at University of Houston.

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Answer-Key---Recombi_36751 - BIOTECHNOLOGY PROBLEM 1 Using...

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