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Biochemistry Techniques in molecular biology 7d5757c6

The enzyme used to "flush" the sticky ends of DNA is

A
Klenow fragment
B
Polynucleotide kinase
C
Alkaline phosphatase
D
Primase
High-Yield Explanation
Flushing of sticky ends means to remove the sticky ends (3' overhangs) for producing blunt-ended DNA (no overhangs). The sticky ends has 3' overhangs which can be digested with Klenow fragment (has 3'-5' exonuclease activity) to produce blunt-end DNA. Tool Role Type II restriction enzymes Cleaves DNA at specific sites DNA ligase Phosphodiester bond formation between 2 fragments Klenow fragments (DNA polymerase I without 5' to 3' exonuclease activity) Used previously for polymerization in PCR reactions Polynucleotide kinase Catalyse the transfer of g phosphate from ATP Alkaline phosphatase Removes the phosphate group from 5' end to prevent self annealing of sticky ends of DNA Reverse transcriptase RNA dependent DNA polymerase for cDNA synthesis RNAase H Removes RNA from DNA-RNA hybrid Exonuclease III Removes nucleotide from 3' end Phage g exonuclease Removes nucleotide from 5' end Terminal nucleotidyl transferase In homopolymer tailing to conve blind end to sticky end

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