Problems with the cloning of foreign DNA in E-coli

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DaVinci
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Problems with the cloning of foreign DNA in E-coli

Post by DaVinci » Sat Aug 20, 2005 1:55 pm

I understand that cloning foreign DNA in Escherichia coli (using a plasmid vector) involves 4 steps, i.e.

1- Cutting the required section of DNA out of the donor DNA and isolating the required piece.
2- Inserting the foreign DNA into a suitable vector.
3- Inserting the recombinant vector into the host cell.
4- Selecting cells that have taken up the recombinant vector.

My question is, what exactly are the problems associated with each of the steps, and how do the techniques used overcome the problems involved? I've read through several books and peer reviewed journals, but haven't found an explaination sound or detailed enough.

Answers would be much appriciated :o

sdekivit
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Post by sdekivit » Sat Aug 20, 2005 4:22 pm

check a book about biochemistry, for example 'Biochemistry' from Stryer.

loupass
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Post by loupass » Wed Sep 21, 2005 9:17 pm

what you need to look up are restriction/modification systems. These are the keys to introducing (or restricting) foreign DNA. Any good molecular biol textbook will provide a good description. Of course, the web is full of potential sites about topics just like this.

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