What is meant by AFLP?
Amplified fragment length polymorphism (AFLP) is a PCR-based technique that uses selective amplification of a subset of digested DNA fragments to generate and compare unique fingerprints for genomes of interest.
What is AFLP in DNA fingerprinting?
The AFLP technique is a powerful DNA fingerprinting technology applicable to any organism without the need for prior sequence knowledge. The protocol involves the selective PCR amplification of restriction fragments of a total digest of genomic DNA, typically obtained with a mix of two restriction enzymes.
What is the difference between RAPD and AFLP?
The AFLP technique is more laborious and time consuming than RAPD methods but is also more reliable, AFLP being able to detect a large number of polymorphic bands in a single lane rather than high levels of polymorphism at each locus such as is the case for SSR methods.
What is AFLP in plant breeding?
AFLP, or amplified fragment length polymorphism, refers to a method where total genomic DNA extracted from plant tissue is digested with restriction enzymes to generate DNA fragments before performing the DNA copying procedure of PCR.
What is the difference between RFLP and AFLP?
Both RFLPs and AFLPs involve differences in the lengths of DNA fragments. The two techniques differ because RFLPs are revealed by digestion with restriction enzymes, whereas AFLPs involve both digestion with restriction enzymes and a selective amplification step.
Why AFLP is a dominant marker?
AFLP are multilocus markers and their mode of inheritance is dominant. The genotyping technology is rather simple. The main advantages of this system are the relative ease of the genotyping, the relative high number of loci detected in each reaction, and the reliability of the system.
What are the steps of AFLP?
This technique involves five major steps, as described in the following sections.
- Step 1: Preparing the AFLP Template. Figure 1.
- Step 2: Ligation Reaction with Restriction Fragments and Adaptors.
- Step 3: Selective PCR Amplification.
- Step 4: Electrophoretic Separation of Amplified DNA Fragments.
- Step 5: Analysis.
What is RFLP and AFLP?
Amplified Fragment Length Polymorphism (AFLP) and Restriction Fragment Length Polymorphism (RFLP) are two such molecular markers (methods) developed in molecular biology to detect genetic variation between organisms. Both methods are equally important and have advantages and disadvantages.
What is AFLP in pregnancy?
AFLP is a rare, but serious, condition of pregnancy in which there is an excessive accumulation of fat in the liver or liver cells. Fat normally accumulates in the liver in the form of triglycerides and fatty acids, but excessive fat can cause liver damage.
Who discovered AFLP?
AFLP-PCR was first described by researcher Pieter Vos and his colleagues in 1995 (Vos et al., 1995). This technique involves five major steps, as described in the following sections.
What are the disadvantages of AFLP?
Disadvantages include the need for purified, high molecular weight DNA, the dominance of alleles, and the possible non-homology of comigrating fragments belonging to different loci.
What is SSR marker?
Microsatellites, otherwise called Simple sequence repeats (Ssrs) or Short Tandem Repeats (Strs), are rehashing sequences of 2-5 base sets of Dna.it is a sort of Variable Number Tandem Repeat (VNTR). Microsatellites are commonly co-prevailing.
What is AFLP marker?
This marker type was named as AFLP or Amplified Fragment Length Polymorphism. AFLP involves the digestion of genomic DNA using restriction endonucleuses, followed by adapter ligation and PCR amplification. The amplified products are visualised on high resolution polyacrylamide gels or automated sequencers.
How are molecular genetic polymorphisms identified with AFLP?
With AFLP, molecular genetic polymorphisms are identified by the presence or absence of DNA fragments following restriction and amplification of genomic DNA. The process consists of four steps: (i) Digestion of genomic DNA
What are the preselective and selective AFLP primers?
For the preselective (PS)-AFLP amplification, one EcoRI and one MseI primer containing a single selective nucleotide. For the selective AFLP amplification, at least one EcoRI and one MseI primer containing a two to three selective nucleotides on each are required.
How do you quantify DNA in AFLP?
The AFLP procedure (1) requires c. 500 ng of genomic DNA (see Note 9) in 5.5 μl volume as starting material. Therefore, proceed to quantify DNA. To obtain the desired concentration the DNA extract can be concentrated in a vacuum oven at 60 °C or diluted using 1x TE buffer.