Discussion 9 Fall 2008 - GENETICS(BIO 325 FALL 2008...

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GENETICS (BIO 325) FALL 2008 Discussion sheet 9 _____________________________________________________________________________ _ 1. Here are the recognition sites of four restriction enzymes: Each enzyme cuts between the bases indicated by the arrows. Which pair of these enzymes will create compatible (complementary) sticky ends? What is the sequence of this compatible sticky end? 2. Restriction endonucleases Bam H1, Nhe I, Xba I, and Eco RI make sequence-specific cuts in DNA at the following sequences: Bam H1 Nhe I Xba I Eco RI 5'-GGATCC-3' 5'-GCTAGC-3' 5'-TCTAGA-3' 5'-GAATTC-3' 3'-CCTAGG-5' 3'-CGATCG-5' 3'-AGATCT-5' 3'-CTTAAG-5' All these enzymes break a phosphodiester bond between the first and second nucleotide of the recognition sequence (counting from the 5' end), leaving a 5' P and a 3' OH. Could ligase covalently join two ends produced by the following enzymes? If so, would ligation regenerate one of the above restriction sites. (1) Two Bam H1 ends? (2) Two Xba I ends? (3) A Bam H1 and an Eco RI end? (4) An Xba I and an Nhe I end?
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