Describe the main components of the ligation reaction and th

Describe the main components of the ligation reaction and their importance in the reaction. What would happen in your ligation reaction if the vector was a closed circular plasmid containing the LacZ gene? What color would the colonies be if you were to transform the ligation described in part a? Why?

Solution

Irrigation reaction required 3 ingredients in addition to Water 2 or more fragments of DNA that have a blunt are compatible cohesive sticky ends and a buffer which contains ATP.

DNA ligase is the enzyme at the heart of DNA ligation reaction it covalent covalently joint the phosphate backbone of DNA with blunt or compatible cohesive ends and its natural role is in repairing double strand breaks in DNA molecule.

The DNA concentration, ligaseconcentration ,temperature, buffer composition all these affect the ligation reaction.

For the insertion of a DNA fragment into a plasmid vector it is preferable to use two different restriction enzymes to digest the DNA so that different ends generated the two different ends can prevent the realisation religation of the vector without any insert and it close the fragment to be inserted in a directional manner.

B. The cell Transformer with vectors containing Recombinant DNA will produce white colonies