GENE CLONING

Recombination

 

• Recombination is a process by which package the DNA of interest into a vector.

• Recombinant DNA = Vector that carry foreign DNA.

• Non-recombinant DNA = Vector that undergo self-ligation and do not carry any foreign DNA.

 

Process:

1. Both of the samples DNA and plasmid DNA (vector) will be restricted by restriction enzyme at restriction      

    recognition site.

• ​Restriction recognition site usually is palindromic sequence.

• After restriction, restricted DNA will produce sticky ends or blunt ends.

• Linker and adaptor will be used when necessary.

 

2. After restriction, both DNA samples will be mixed and incubate with the presence of ligase.

• Different ligase will be used depend on the cutting pattern

 

3. Ligation between the DNA samples occurs.

• Possible results:

  • Recombination success: Ligation between vector DNA and DNA of interest.

  • Recombination failed: Self- ligation of vector occur.