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Random Amplified Polymorphic DNA - RAPD - The technical complexity of RFLP analysis, coupled with the widespread use of short lived radioisotopes in the detection method, questions the suit­ability of routine use of RFLPs in large scale use.

Williams et al. (1990) described a DNA polymorphism assay based on the polymerase chain reaction (PCR) amplified random segments with single primers of arbitrary nucleotide sequence.

They suggested that these polymorphisms might be called RAPD markers, after random amplified polymorphic DNA.

The RAPD method is technically simple, quick to perform, requires small amounts of DNA and involves no radioactivity; it is well suited for uses in large sample throughout systems required for breeding, population genetics and biodiversity.

Rapid amplification of polymorphic DNA method is proving to be a very  promising technique for genetic analysis using RFLPs.

In this technique the polymorphic probes are sequenced and oligonucleotide synthesis carried out to synthesize fragments based on the sequence information revealed.

These fragments are then utilized as primers for DNA amplification by the PCR technique.

This results in the production of a sufficient amount of DNA fragments downstream of the primers, which can be detected on an agarose gel after staining with ethidium bromide.

After the PCR step, the DNA samples are cut by appropriate restriction enzymes and separated on agarose gels.

They are then stained with ethidium bromide to detect the amplified fragments. These reveal an RFLP pattern which can be utilized for further analysis.

This method obviates the radioactive method and furthermore, is much faster than the existing method of screening by Southern hybridization.

All in all, RAPD seems to be a very promising method and can be used hand in hand with the existing RFLP technique.

 
 

 
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