Cloning 101

newsnet customer customer at newsnet.com
Tue Nov 29 04:17:27 EST 2005


Hi,

Whats is the general process of cloning a gene into a vector? Heres what I do but I dont think its efficient - still a newbie.

1. first thing i look at is where I am cloning into the vector. I try to find two unique restriction sites.
2. I then check if the ends of my gene when cut will be compatible to the restriction sites when cut. Here is where i find myself drawing A, T, G, and C and overhangs and seeing if it fits all together. This will almost never happen. So i goto step 3.
3. Design primers which will include compatible restriction sites to the restriction sites in the vector and amplify my gene.

Question:
Is there a quick way of knowing which restriction sites should be included in the primers so that it will be compatible to the sites in the vector.
I'm just guessing:

Gene
BamH1 ... EcoR1             

goes into: 

Vector
BamH1...EcoR1

Is there some sort of rule/pattern?

Cheers
NC

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