E. Which enzymes listed below could be used to repair (generate the ends that wi

Question

E. Which enzymes listed below could be used to repair (generate the ends that will be usable by DNA ligase) the DNA ends listed below. If more than one enzyme is required, indicate if they are used together or sequentially. If the latter specify the order. Some enzymes may be used multiple times: others may not be used at all. Enzymes: DNA polymerase I Klenow fragment (polymerization domain only) .Polynucleotide kinase (specific for 5 OH groups) Mung bean nuclease (specific for single-stranded DNA overhangs. It leaves 5" phosphate or 3 OH: not effective against short 3'-phosphate overhangs.) .Shrimp alkaline phosphatase acts on 5' and 3' phosphates, recessed or protruding Other nucleases described by New England Biolabs at https://www.neb.com/products/dna-modifying-enzymes-and-cloning-technologies/nucleases/nucleases Ends: OH(3) OH(3) OPO3 (5') OPO3(5') OH(3') OPO3(5') -OH(3) OH(5) OPO3(3') OH(5) OH(3') OH(5')

Explanation / Answer

Ans)

i. DNA Polymerase I klenow fragment (polymerization domain only) :

________________________OH (3')

________________________OH (5')

ii. Polynucleotide Kinase (specific 5' OH groups) :

_________________________OH (3')

_________________________OPO3 (5')

iii. Mung bean nuclease:

___________OH (3')

_____________________OH (5')

iv. shirmp alkaline phosphatase

___________________OH (3,)

_________________________OPO3 (5')

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