Linkers- Short stretches of dsDNA of known nucloetide sequence (length 8-14 bp )and have a recognition site for 3-8 restriction enzymes.
Ligated to blunt ends of DNA by ligase.Because of the high concentration of these small molecules present in the reaction, the ligation is efficient when compared with blunt-end ligation of large molecules. The cohesive ends are generated by digesting the DNA with appropriate RE that generates cohesive ends by cleaving in the linkers. The problem with linker is that the sites for the enzyme used to generate cohesive ends may be present in the target DNA fragment. This drawback limits the use of linkers for cloning.
Adapters are linkers with cohesive ends or a linker digested with RE, before ligation.It already has one sticky end.The most widely used definition is cut linkers also called as adapters.
They are not perfectly double stranded non single stranded. By adding adaptors to the ends of a DNA, sequences that are blunt can be converted into cohesive ends and used for cloning .
Ligated to blunt ends of DNA by ligase.Because of the high concentration of these small molecules present in the reaction, the ligation is efficient when compared with blunt-end ligation of large molecules. The cohesive ends are generated by digesting the DNA with appropriate RE that generates cohesive ends by cleaving in the linkers. The problem with linker is that the sites for the enzyme used to generate cohesive ends may be present in the target DNA fragment. This drawback limits the use of linkers for cloning.
Adapters are linkers with cohesive ends or a linker digested with RE, before ligation.It already has one sticky end.The most widely used definition is cut linkers also called as adapters.
They are not perfectly double stranded non single stranded. By adding adaptors to the ends of a DNA, sequences that are blunt can be converted into cohesive ends and used for cloning .
Sticky ends are desirable for DNA cloning , but sometimes blunt ends are present .Blunt ends will binds to the blunt ends of target to form sticky ends.
Image source-http://www.slideshare.net/drsandeepaiims/gene-cloning-sandeep-jan-2016
Thank you for visiting my blog.Please feel free to share your comment on this article ,Please subscribe and share the articles to get more such articles.
No comments:
Post a Comment