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6 years ago in Cloning Method By Rinku

Restriction enzyme vs gateway recombination cloning method

Hello all. Should I use restriction enzyme or gateway recombination cloning method for my study? Which is more reliable?

All Answers (5 Answers In All)

By Vinod D Answered 6 years ago

Rinku the type of cloning method to be used more often depends on your cloning study. However, restriction enzyme or endonuclease based cloning is often considered as the classic cloning approach and is most used by the research students.

By Novi Tulipe Answered 6 years ago

Gateway cloning is based on recombination cloning approach. The advantage of using this method is that the reaction which moves a DNA piece from one plasmid into another is conducted through a single recombination reaction. To utilise this method, the fragment of DNA you like to clone should be surrounded by certain recombination sites. Once the DNA fragment is cloned into a donor plasmid, it is shuttled into suitable gateway destination vector.

By Rinku Answered 6 years ago

I want to chop genomic DNA into tiny pieces to prepare a library. Can I use gateway recombination method for the same?

By Nadeemimi Answered 6 years ago

No, for cutting genomic DNA into smaller pieces you have to use restriction enzyme cloning method. To know more about this method visit https://www.khanacademy.org/science/biology/biotech-dna-technology/dna-cloning-tutorial/a/restriction-enzymes-dna-ligase.

By Keerthi Gupta Answered 6 years ago

https://www.embl.de/pepcore/pepcore_services/cloning/cloning_methods/restriction_enzymes/ this is a good website. It explains you vector preparation, insert preparation and ligation methods i.e the steps involved in methylation of DNA as well.

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