KpnI is a restriction endonuclease that is used in molecular biology applications to cleave the DNA recognition site 5′-GGTAC/C-3′, resulting in DNA fragments with 3′-cohesive termini. This type II restriction enzyme is isolated from the bacterium Klebsiella pneumoniae. KpnI produces a “sticky” end cleavage, generating a four-base 3′ overhang. This overhang can be valuable in constructing recombinant DNA by facilitating the joining of complementary DNA fragments.
Applications:
- Molecular cloning: Kpn I is commonly used in molecular cloning techniques to insert genes into plasmids.
- Restriction site mapping: Kpn I can be used to map restriction sites on DNA molecules.
- Genotyping: Kpn I can be used for genotyping studies to identify genetic variations.
- Southern blotting: Kpn I can be used in Southern blotting techniques to detect specific DNA sequences.
- Restriction fragment length polymorphism (RFLP): Kpn I can be used in RFLP analysis to study genetic variations.
- SNP analysis: Kpn I can be used to analyze single nucleotide polymorphisms (SNPs).
