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Sticky end & blunt end ligation
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#Ligation
The concept is used in molecular biology, especially in cloning or when sub cloning inserts DNA into vector DNA. Such ends may be generated by restriction enzymes that cut the DNA – a staggered cut generates two sticky ends, while a straight cut generates blunt ends.
DNA ligases repair nicks in double stranded DNA with 3’ OH and 5’ phosphate ends. Typically, bacterial ligases are NADH-dependent, while eukaryotic ligases are ATP-dependent; however, despite T4 DNA ligase’s existence being confined within a prokaryote, it encodes an ATP-dependent ligase.
T4 DNA ligase has a lower affinity for DNA than multidomain ligases, which could also explain its sensitivity to salt concentration. DNA ligation occurs in a three-step reaction. For more videos please click the following links
#Ligation
The concept is used in molecular biology, especially in cloning or when sub cloning inserts DNA into vector DNA. Such ends may be generated by restriction enzymes that cut the DNA – a staggered cut generates two sticky ends, while a straight cut generates blunt ends.
DNA ligases repair nicks in double stranded DNA with 3’ OH and 5’ phosphate ends. Typically, bacterial ligases are NADH-dependent, while eukaryotic ligases are ATP-dependent; however, despite T4 DNA ligase’s existence being confined within a prokaryote, it encodes an ATP-dependent ligase.
T4 DNA ligase has a lower affinity for DNA than multidomain ligases, which could also explain its sensitivity to salt concentration. DNA ligation occurs in a three-step reaction. For more videos please click the following links
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