Gene and Protein Engineering

Nucleic acid hybridization uses single stranded nucleic acid molecules to form double-stranded molecules by interacting sets of single stranded molecules that have a high base complementarity. It involves a labelled nucleic acid probe identify the related DNA/RNA.

DNA probes can either be cell based DNA cloned or cloned through PCR. They are labelled by incorporating labelled dNTPs in an in vitro DNA synthesis reaction.

RNA probes are generated from DNA that has been cloned in a specialized plasmid vector that contains a phage promoter sequence next to the multiple cloning sites.

Oligonucleotide probes are short single stranded pieces of DNA made by adding mononucleotides together from the 3'end and are labelled with 32P atom at the 5' end.

Labelling of DNA in vitro is done by nick-translation, random primed labelling or PCR mediated labelling.

The denaturation of the double stranded DNA probe occurs through heating the solution so the H bonds are disrupted. This temperature is dependent on strand length, base composition and chemical environment.

Mammalian genomes with a base composition of 40% GC will denature…


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