1. Traditional Gibson Gateway
Cloning Strategies
Traditional
Gibson
Gateway

2. Traditional cloning Time consuming Cheaper than other methods
Limited with restriction enzymes
Lots of screening and sequencing
Start over from beginning each time you clone it into a new vector.

3. Gibson Cloning

4. Limitations- termini of DNA sequence should not have stable single strand DNA secondary structure(ie hairpin loop) as it would interfer with the annealing step.
Limit of 2-3kb per piece of DNA
Still need to sequence after cloning is finished.

5. Gateway Cloning

6. Gateway Cloning

7. Fast, one hour, room temperature cloning reactions with >99% efficiency deliver the clone you need
Maintaining orientation and reading frame without using restriction enzymes or ligation makes expression-ready clones
Eliminating re-sequencing ensures consistent results throughout your experiment using the same clone from target identification to validation
Shuttling insert DNA from one expression vector to another affords flexibility while simplifying your cloning workflow

8. Limitations-Does not save time or money over traditional cloning.
Only useful if you are planning on doing many things with a gene of interest.
Not any better for screening genes than traditional cloning.