Question 2: Sticky Ends 4 points Imagine you want to ligate (join) two separate

Question

Question 2: Sticky Ends 4 points Imagine you want to ligate (join) two separate pieces of DNA together so that they form the sequence shown in the middle. One method you could use is blunt end ligation between the two pieces of completely double-stranded (ds) DNA. However, that will also lead to many products that you do not want. Therefore, you decide to use a sticky-end design. Remember, "sticky-end" refers to any single-stranded DNA that protrudes from the ends of a fragment of double-stranded DNA. Write the sequence of two pieces of DNA that will anneal by sticky end igation to make the sequence shown below. Design your DNA to have sticky ends that are each 4 nt long, HINT: start by drawing your design on the DNA sequence shown and then write your answer below, showing the two pieces of DNA with appropriate sticky ends. There are several correct answers possible.

Explanation / Answer

Restriction endonucleases are enzymes that recognize a specific DNA sequence, called a restriction site, and cleave the DNA within or adjacent to that site. For example, the restriction endonuclease EcoR I AATTC, many of them are palindromic. But also, are enzymes that recognize non-palindromic sequences. For that reason, we can choose the restriction site and use a non-palindromic restriction endonuclease.

The following two sequences could be one possible solution.

5´-GGA                        TTGTGCGA-3´

3’-CCTAACA                         CGCT-5’

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