What is PCR and RAPD?
Random amplified polymorphic DNA (RAPD) is a PCR based technique for identifying genetic variation. It involves the use of a single arbitrary primer in a PCR reaction, resulting in the amplification of many discrete DNA products. Such polymorphisms thus behave as dominant genetic markers.
Why RAPD is dominant marker?
On average, each primer directs amplification of several discrete loci in the genome so that allelism is not distinguishable in RAPD patterns. In other words, it is not possible to distinguish whether a Page 4 DNA segment is amplified from a locus that is heterozygous or homozygous. RAPD markers are therefore dominant.
Is RAPD a molecular marker?
markers are DNA fragments from PCR amplification of random segments of genomic DNA with single primer of arbitrary nucleotide sequence.
Why RAPD is done?
RAPD technology provides a quick and efficient screen for DNA sequence based polymorphism at a very large number of loci. The major advantage of RAPD includes that, it does not require pre-sequencing of DNA. The vast range of potential primers that can be used, give the technique great diagnostic power.
What are the optimized conditions for RAPD-PCR protocol?
The optimized conditions for RAPD protocol are given in Table 1. The size of the amplified fragments ranged from 5000-2000 bp. Gel fragmentation of RAPD-PCR amplification product of plant genomic DNA 50 ng. Lane 1-3 represents RAPD-PCR products amplified from genomic DNA of study plants.
What is the difference between PCR and RAPD?
Unlike traditional PCR analysis, RAPD (pronounced “rapid”) does not require any specific knowledge of the DNA sequence of the target organism: the identical 10-mer primers will or will not amplify a segment of DNA, depending on positions that are complementary to the primers’ sequence.
Are co-dominant RAPD markers detected in PCR?
Co-dominant RAPD markers, observed as different-sized DNA segments amplified from the same locus, are detected only rarely. PCR is an enzymatic reaction, therefore the quality and concentration of template DNA, concentrations of PCR components, and the PCR cycling conditions may greatly influence the outcome.
What is polymorphic RAPD marker in PCR?
The polymorphic RAPD marker band is isolated from the gel. It is amplified in the PCR reaction. The PCR product is cloned and sequenced. New longer and specific primers are designed for the DNA sequence, which is called the Sequenced Characterized Amplified Region Marker (SCAR).