In cloning,how do the DNA fragments get inserted int a plasmid?

PDG · #1 (**permalink**) 09-14-2009, 07:16 PM

Plasmids are usually treated with alkaline phosphatase to prevent self-ligation.In such a case,can the inserted fragment be incoporated into plasmid in more than two orientations?

wangleipurdue · #2 (**permalink**) 09-24-2009, 07:37 PM

If you digest the fragment by enzymes that give you sticky ends,plus the two enzyme sites you use are different, normally,the fragment should just insert in one direction. If the two sites you choose are the same (it is very rare), or you use two sites give you blunt ends, the fragment can be inserted in two direction.

you can get information of enzymes at [Only registered users see links. ]

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In cloning,how do the DNA fragments get inserted int a plasmid?

In cloning,how do the DNA fragments get inserted int a plasmid? - DNA Techniques

In cloning,how do the DNA fragments get inserted int a plasmid? - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.