Strategies of genetic engineering - Tricks with DNA primers

6 important questions on Strategies of genetic engineering - Tricks with DNA primers

Can you join a DNA sticky end with a DNA blunt end?

No.

What is the barrier to prevent the eukaryotic genomic clones/genes from being expressed in bacteria?

Eukaryotic genes contain introns.

What are conventional cloning methods?

Cleavage using restriction enzymes.
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What is nonconventional cloning?

Recombination.

What is TA cloning?

A subcloning technique that avoids the use of restriction enzymes and is easier and quicker than traditional subcloning.
It takes advantage of the 3′ A overhangs added during amplification. These sticky ends enable insertion into a linearized vector with complementary 3′ T overhangs.

TA cloning is used for preparation of gene library. You need to have specific plasmid for insertion.
Taq polymerase adds to  3´ prime deoxyadenosine

What is GC cloning?

The GC Cloning technology is based on the observation that nonproofreading DNA polymerases add a single 3´-G to DNA molecules, either during PCR or as a separate G-tailing reaction to any blunt DNA.

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