Random amplified polymorphic dna rapd pdf

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RAPD 1. Presented by: Abhi Giri M. Jayani M. It works. Hence a popular method. How it Works? If another DNA template genome B was obtained from a different yet related source, there would probably be some differences in the DNA sequence of the two templates. Suppose there was a change in sequence at primer annealing site 2 Figure 2 genome A Figure 3 genome B 9. Find those sequences which have just enough variation to allow us to detect differences among the organisms that we are studying.

The scoring can be done based on the banding profiles which is clear and transparent Fig. Template DNA 3. The DNA of a selected species is isolated. An excess of selected decaoligonucleotide added. During this process, the decaoligonucleotide will pair with the homologous sequence present at different locations in the DNA.

DNA replication extend the decaoligonucleotide and copy the sequence continuous with the sequence with which the selected oligonucleotide has paired. Amplification will takes place only of those regions of the genome that has the sequence complementary to the decaoligonucleotide at their both ends. After several cycles of amplification the DNA is subjected to gel electrophoresis.

The amplified DNA will form a distinct band. Complementary strand synthesis 35 to 45 cycles Amplified products separated by gel electrophoresis Bands detected by Ethidium bromide staining The PCR product is cloned and sequenced.

Probe Reagents for Functional Genomics. How It Works Unlike traditional PCR analysis, RAPD pronounced "rapid" does not require any specific knowledge of the DNA sequence of the target organism: the identical mer primers will or will not amplify a segment of DNA, depending on positions that are complementary to the primers' sequence. Example RAPD is an inexpensive yet powerful typing method for many bacterial species.

Thus, the RAPD technique is notoriously laboratory dependent and needs carefully developed laboratory protocols to be reproducible. Mismatches between the primer and the template may result in the total absence of PCR product as well as in a merely decreased amount of the product.

Thus, the RAPD results can be difficult to interpret. It is amplified in the PCR reaction. The PCR product is cloned and sequenced. You are here: NCBI. External link. Please review our privacy policy.