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what is digestion with restriction enzymes?

what is digestion with restriction enzymes? - DNA Techniques

what is digestion with restriction enzymes? - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


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Old 06-11-2009, 03:17 AM
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Default what is digestion with restriction enzymes?



Im an intern working with plant pathology and I recently did this lab thing called Genejet Plasmid Miniprep Kit and then digestion with restriction enzymes. And I had absolutely no idea what any of it is about. I dont understand what it is Im doing. sooooo if someone could explain the concept and what the digestion part was/how to do it in like a really simple-to-understand way that would be great thanks
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Old 06-11-2009, 06:45 AM
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Default Re: what is digestion with restriction enzymes?

Basically, miniprep is short for minipreparation. It is a procedure which includes you culturing a bacteria culture overnight to get a lot of bacteria that can be centrifuged the next day to obtain a pellet. What miniprep does is to lyse the bacteria so that the plasmids can be isolated. This involves the usage of a few buffers that breaks the membrane of the bacteria, washing the DNA and eluting the plasmids.
The plasmids can be used for restriction digestion to check for inserts that you need or for you to insert DNA inserts into it. Basically, you make use of restriction enzymes, which recognizes restriction sites on the plasmid and cut them up. The same or different restriction enzymes can be used on another DNA which you want to insert into the plasmid. Ligation is then performed to ligate the DNA insert into the plasmid.
I hope this is clear enough for you? You can look up in google for more details. Or else, provide us with more details on what you do so it will be easier for us to tell you what is going on. ALternatively, you can always ask your supervisor.
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Old 06-11-2009, 06:46 AM
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Default Re: what is digestion with restriction enzymes?

Basically, miniprep is short for minipreparation. It is a procedure which includes you culturing a bacteria culture overnight to get a lot of bacteria that can be centrifuged the next day to obtain a pellet. What miniprep does is to lyse the bacteria so that the plasmids can be isolated. This involves the usage of a few buffers that breaks the membrane of the bacteria, washing the DNA and eluting the plasmids.
The plasmids can be used for restriction digestion to check for inserts that you need or for you to insert DNA inserts into it. Basically, you make use of restriction enzymes, which recognizes restriction sites on the plasmid and cut them up. The same or different restriction enzymes can be used on another DNA which you want to insert into the plasmid. Ligation is then performed to ligate the DNA insert into the plasmid.
I hope this is clear enough for you? You can look up in google for more details. Or else, provide us with more details on what you do so it will be easier for us to tell you what is going on. ALternatively, you can always ask your supervisor.
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