What is PCR and RAPD?
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. The technique was developed independently by two different laboratories (Williams et.
What is Lactobacillus species by PCR?
Lactobacillus species are used as probiotics and play an important role in fermented food production. However, use of 16S rRNA gene sequences as standard markers for the differentiation of Lactobacillus species offers a very limited scope, as several species of Lactobacillus share similar 16S rRNA gene sequences.
What is the purpose of RAPD?
RAPD is involved in the amplification of DNA by PCR using arbitrary short primer. These short sequence primers can be used for any genetic material of an organism even with an unknown target sequence.
What type of product is generated after RAPD?
Developing Locus-specific, Co-Dominant Markers from RAPDs 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).
How do you do RAPD PCR?
The RAPD-PCR protocol consisted of an initial denaturing step of 2 min at 95 °C, followed by 45 cycles at 95 °C for 1 min (denaturation), 36 °C for 1 min (annealing of primers), and 72 °C for 2 min (extension). Cycling was concluded with a final extension at 72 °C for 4 min, and then held indefinitely at 4 °C.
What are the advantages of RAPD?
Main advantages of the RAPD technology include (i) suitability for work on anonymous genomes, (ii) applicability to problems where only limited quantities of DNA are available, (iii) efficiency and low expense.
What are the drawbacks of RAPD PCR?
The disadvantage of this method is the poor reproducibility of fingerprints, and it requires strict standardisation of PCR conditions considering that the utilisation of different concentrations of DNA polymerases, DNA template and primer ratios or annealing temperatures can lead to differences in the final results ( …
Is RAPD a DNA sequencing based method?
RAPD is a PCR based technology in which DNA polymorphism assay is based on the amplification of random DNA segments with single primers of the arbitrary nucleotide sequence. A single type of primer is used to anneals to the genomic DNA at two different sites on complementary strands.
Why do I have Lactobacillus overgrowth?
Frequently, a woman may present with chronic vaginal discharge that has been treated with a variety of antifungals and antibiotics. These treatments can provoke a change in the vaginal pH leading to an overgrowth of lactobacilli. Other triggers may include sensitivity to soaps, wipes, pads, lubricants, etc.
What is Lactobacillus infection?
Lactobacillus organisms are rarely associated with pathology in immunocompetent people, but in the presence of risk factors and underlying conditions, they can cause infections such as endocarditis, bacteremia, neonatal meningitis, dental caries, and intra-abdominal abscesses including liver abscess, pancreatic …
How many primers are used in RAPD?
Then RAPD is employed using two different primers to produce a banding pattern that is then assessed by agarose gel electrophoresis.
What is the difference between RAPD and AFLP?
Key: RAPD = random amplified polymorphic DNA; RFLP = restriction fragment length polymorphism; AFLP = amplified fragment length poly- morphism; SSR = simple sequence repeat. from this population (Gerage et al., 1988, 1989).
What is the disadvantage of RAPD or RFLP in laboratory?
3) RAPDs are simple, quick, and cost effective compared to RFLP [17, 18]. However, RAPDs also have some disadvantages, these include 1) the repeatability and reliability of RAPD polymorphic profiles are poor . 2) Some non-specific and therefore non-reproducible binding of primers occurs.