Dus: Sun, Oct 19
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67
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juestion 7 of 19
Stacked
_(_())_(_())
Suppose you can synthesize a short duplex DNA fragment with any sequence you desire. With this synthetic fragment and the procedures described in parts
A through
G, you could remove an EcoRI restriction site from a DNA molecule and incorporate a new BamHI restriction site at approximately the same location.
There are at least two different methods for replacing an EcoRI restriction site with a BamHI restriction site.
Complete the passage describing the method that inserts a DNA fragment with blunt ends.
Cut the DNA with
_(_())_(_()) then treat the DNA with
_(_())_(_()) to create blunt ends.
Correct Answer
Correct Answer
Then ligate a synthetic fragment with the
_(_())_(_()) recognition sequence between the two resulting blunt ends.
Correct Answer
A second method would be to synthesize a DNA fragment that would ligate efficiently to the sticky ends generated by EcoRI cleavage and introduce a BamHI cleavage site, without regenerating the EcoRI cleavage site.
Select the synthetic DNA fragment you could use to replace an EcoRI restriction site with a BamHI restriction site.
O
5^(')- GATCGAATTC
-3^(')
3^(')-CTTAAGCTAG-5'
◻
5'-TTAAGGATCC-3'
3^(')-CCTAGGAATT-5'
◻
5^(')- AATTGGATCC
-3^(')
3^(')-CCTAGGTTAA-5*
◻
5^(')-AATTCCTAGG-3'
3'-GGATCCTTAA-5'
vicomed Answer