DNA Ligation — Transcript
Full transcript
- 0:01DNA ligation is the formation of a
- 0:04covalent bond between adjacent DNA
- 0:07fragments frequently a vector and a gene
- 0:10of interest ligation is usually the
- 0:14final step before transformation in the
- 0:17cloning workflow
- 0:20DNA ligase may be used to join
- 0:23double-stranded DNA fragments with
- 0:25either blunt or cohesive ends to form
- 0:27recombinant DNA plasmids
- 0:31a DNA ligase catalyzes the formation of
- 0:35a phosphodiester bond between juxtaposed
- 0:37five prime phosphate and three prime
- 0:40hydroxyl termini in duplex DNA ligation
- 0:44proceeds in three steps
- 0:47initially the ligase is self ademma
- 0:50lated by reaction with free ATP or nad
- 0:55next via dental group is transferred to
- 0:59the five prime phosphoryl aided end of
- 1:01the donor strand
- 1:04lastly the formation of the
- 1:06phosphodiester bond proceeds through the
- 1:09reaction of the adenylate a donor end
- 1:11with the adjacent 3 prime hydroxyl
- 1:13acceptor accompanied by the release of a
- 1:16MP
- 1:20while t4 DNA ligase is the most commonly
- 1:23used lacus for cloning there are several
- 1:26options from any be to choose from
- 1:29including convenient master mix formats
- 1:32for help with selection of a DNA ligase
- 1:36optimized for a specific end type and
- 1:38application view our selection chart at
- 1:41any be stick together com visit clone
- 1:46with any be com for a full list of
- 1:49products available for cloning
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