Linkers: Joining molecules, Cloning of Blunt DNA Fragments/ cDNA molecules

Linkers are chemically synthesized double standard DNA oligonucleotides with an internal restriction cleavage site and blunt ends.
Linkers can be put on blunt ended DNA fragments and cDNA molecules to generate cohesive ends after digestion with a particular restriction endonuclease.
Linkers have 5’ PO4 and 3’OH like a natural DNA
Ligation of linkers with DNA fragment is quite easy by providing an excess concentration of chemically synthesized linkers, even though it is blunt end ligation.
During the ligation, linkers can bind to each other and also they can bind to linker which has bound to blunt end DNA molecule and forms a chain like structure
If the DNA to be cloned has the same restriction site internally as the restriction site in a vector, then the linkers with particular restriction site can not be used.

[@rashmipandey7425]

Sir I have a question, while making phagemid vector do we insert ss DNA fragment of M13 into the plasmid

[@Dr.DNA-Primer]

No, double standard M13 fragment will be inserted. M13 genome converted into double standard that is called replicative form. Those replicative forms can be isolated.

[@Dr.DNA-Primer]

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[@rashmipandey7425]

Thanks a lot sir 🙏 I was really confused about this

[@Dr.DNA-Primer]

You are most welcome,