You are tasked with cutting a piece of DNA with the following two restriction en

Question

You are tasked with cutting a piece of DNA with the following two restriction enzymes in the same reaction

EcoR1

5' G"AATTC 3'

3' CTTAA"G 3'

Sma1

5' CCC"GGG 3'

3' GGG"CCC 5'

5'-CTTAGCGCTTCCCGGGCTCCCACTGAATTCGCCAATATGATCAG-3'

3'-GAATCGCGAAGGGCCCGAGGTGACTTAAGCGGTTATACTAGTC-5'

How many restriction fragments would you obtain? Please write out the fragments paying particular attention to what would the ends of each piece of DNA look like. Please comment on whether each end would be sticky or blunt.

What I am confused about on this question is how to write out the restriction fragments and how to know whether each end is sticky or blunt. I believe there are 6 total restriction fragments.

Explanation / Answer

see, on digestion of DNA with Eco R1 we get these fragments

5'-CTTAGCGCTTCCCGGGCTCCCACTG                 and        AATTCGCCAATATGATCAG-3'

3'-GAATCGCGAAGGGCCCGAGGTGACTTAA                              GCGGTTATACTAGTC-5'

these fragments are with sticky ends.

on digestion with Sma 1, we get

fragment 1                                  fragment 2                                         fragment3

5'-CTTAGCGCTTCCC(blunt end)    and (blunt) 5`-GGGCTCCCACTG (sticky)     and   (sticky) AATTCGCCAATATGATCAG-3'(blunt

3'-GAATCGCGAAGGG(blunt end)         (blunt) 3`-CCCGAGGTGACTTAA(sticky)            (sticky) GCGGTTATACTAGTC-5'blunt

so on both digestions, 3 fragments will appear.

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