Table of contents | |
Introduction | |
Steps of AFLP | |
Applications | |
Advantages of AFLP marker | |
Disadvantages of AFLP | |
Summary |
Markers are the tools that are used to distinguish DNA, individuals, populations or species. All the molecular marker techniques fall under two categories -
The Example of restriction based technique is RFLP marker, In this technique the DNA is restricted with specific Restriction Endonucleases.
PCR-based markers include SSRs, RAPDs, ILPs etc.
In 1995 Peter Vos et al. developed a new marker type which is the combination of both the restriction based as well as the PCR based method. This marker type was named as AFLP or Amplified Fragment Length Polymorphism.
AFLP technique requires very good quality of intake and pure genomic DNA, which should be free from protein and contaminants.
The next step is the digestion with restriction endonucleuses. A rear cutter such as Eco R1 and a frequent cutter such as Mse-1 is used producing sticky ends.
Adapters are double-stranded short oligonucleotide sequences of usually 14 to 20 base pairs. Two different adapters are used one each for Eco R1 and another for Mse1.
These adapters of known sequences serves as the target for PCR amplification.
(a) Pre-selective amplification
It is the first round of amplification, in which few fragments are selectively amplified.
A PCR reaction is set which contains
The genomic DNA,
(b) Selective Amplification
A second round of amplification is known as a selective amplification. In selective amplification more stringent primers (primer contain additional 3 nucleotides) are used to reduce the number of fragments amplified.
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1. What is AFLP? |
2. What are the steps involved in AFLP? |
3. What are the applications of AFLP? |
4. What are the advantages of AFLP markers? |
5. What are the disadvantages of AFLP? |
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