MCB 150 Frequently Asked Questions

Genetic Engineering and Recombinant DNA Technology

I am really confused about the second step of finding the cell clones that are carrying the insert we're looking for (aka screening). What's its purpose?

Lots of cells can be transformed and survive the selection process. But many, if not most, of them will simply be cells that took in a plasmid vector that religated back on itself instead of a plasmid vector that actually got a piece of DNA ligated into it. The former we don't care about, the latter we do. The screen allows us to quickly differentiate between the two, based on the presence or absence of reporter gene expression.

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