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Sticky ends can be converted to blunt ends by a process known as blunting, which involves filling in the sticky end with complementary nucleotides. This yields a blunt end, however, sticky ends are often preferable, meaning the main use of this method is to label DNA by using radiolabeled nucleotides to fill the gap. [4]
Blunt-end ligation, however, is much less efficient than sticky end ligation, typically the reaction is 100X slower than sticky-end ligation. Since blunt-end does not have protruding ends, the ligation reaction depends on random collisions between the blunt-ends and is consequently much less efficient.
EcoRI creates 4 nucleotide sticky ends with 5' end overhangs of AATT. The nucleic acid recognition sequence where the enzyme cuts is G↓AATTC, which has a palindromic complementary sequence of CTTAA↓G. [2] Other restriction enzymes, depending on their cut sites, can also leave 3' overhangs or blunt ends with no overhangs.
Sticky ends of DNA however are more likely to successfully bind with the help of a DNA ligase because of the exposed and unpaired nucleotides. For example, a sticky end trailing with AATTG is more likely to bind with a ligase than a blunt end where both the 5' and 3' DNA strands are paired.
EcoRI digestion produces "sticky" ends, whereas SmaI restriction enzyme cleavage produces "blunt" ends: Recognition sequences in DNA differ for each restriction enzyme, producing differences in the length, sequence and strand orientation (5' end or 3' end) of a sticky-end "overhang" of an enzyme restriction. [31]
Polymerases (such as Phusion) or restriction enzymes that produce blunt ends can also be used for TOPO cloning. Rather than relying on sticky ends, the blunt end TOPO vector has blunt ends where the topoisomerase molecules are bound. Commercial kits, such as the Zero Blunt® Cloning Kit from Invitrogen, are available. [2]
The Sticky’s first season finale on Prime Video opens with the trio stuffing Bo’s body into a deep freezer, only she had already alerted Mike’s Boston crime family about his goings-on in Canada.
Most restriction endonucleases cleave the DNA strand unevenly, leaving complementary single-stranded ends. These ends can reconnect through hybridization and are termed "sticky ends". Once paired, the phosphodiester bonds of the fragments can be joined by DNA ligase. There are hundreds of restriction endonucleases known, each attacking a ...