When you transform a PCR product into a plasmid with a certa

When you transform a PCR product into a plasmid with a certain restriction enzyme, what is the purpose of cleaving the transformed plasmid with different restriction enzymes ?

Solution

Main purpose of cleaving plasmid with different restriction enzymes:

Restriction enzymes are belongs to a larger class of enzymes called nucleases. These are 2 kinds’ exonucleases and endonuclease.

Exo-nucleases remove nucleotides from the ends of the DNA whereas endonucleases makes cut at specific portion within the DNA.

Restriction endonucleases enzymes always cut DNA molecules at a particular point by recognizing a specific sequence of 6 base pairs. This specific base sequence is known as recognition sequence.

There are more than 900 Restriction endonucleases enzymes are isolated from 230 strains of bacteria. Each of which recognize different recognition sequence.

Each Restriction endonucleases enzyme functions by inspecting the length of DNA sequence & recognizes a specific palindromic nucleotide sequences in DNA.

Each Restriction endonucleases enzyme differs for sequence Recognition in DNA. These Restriction endonucleases enzyme mainly produce difference in the sequence, length and orientation of strands.