Understanding Amplified Fragment Length Polymorphism (AFLP)
Which one of the following statements is correct for amplified-fragment length polymorphism (AFLP) ?
(1) PCR using a combination of random and gene- specific primers.
(2) PCR amplification followed by digestion with restriction enzymes.
(3) Digestion of DNA with restriction enzymes followed by one PCR step.
(4) Digestion of DNA with restriction enzymes followed by two PCR steps.
Amplified Fragment Length Polymorphism (AFLP) is a widely used molecular marker technique in genetics and molecular biology. It is based on the selective amplification of DNA fragments produced by digestion with restriction enzymes. AFLP is known for its high reproducibility and sensitivity, making it useful for genome mapping, population genetics, and evolutionary biology studies.
Correct Answer: (4) Digestion of DNA with restriction enzymes followed by two PCR steps
The correct statement is (4) Digestion of DNA with restriction enzymes followed by two PCR steps. AFLP involves an initial digestion of genomic DNA using restriction enzymes, followed by selective amplification using two PCR steps.
What is AFLP?
AFLP is a technique used to detect DNA polymorphisms by cutting genomic DNA with restriction enzymes and amplifying the fragments using PCR. It allows for the detection of genetic variation at a high resolution without prior knowledge of the genome sequence.
Keyphrase: Amplified Fragment Length Polymorphism (AFLP)
Amplified Fragment Length Polymorphism (AFLP) is a molecular technique that combines restriction enzyme digestion and PCR amplification to identify genetic variation.
Steps Involved in AFLP
1. Restriction Enzyme Digestion
- Genomic DNA is digested with two different restriction enzymes:
- A rare cutter (e.g., EcoRI)
- A frequent cutter (e.g., MseI)
2. Adapter Ligation
- Short, double-stranded adapters are ligated to the sticky ends produced by restriction digestion.
- Adapters provide primer-binding sites for PCR.
3. Pre-Selective PCR Amplification
- A PCR is performed using primers complementary to the adapter sequences.
- The primers contain one or two additional nucleotides at the 3′ end to enhance specificity.
4. Selective PCR Amplification
- A second PCR is performed using primers with additional selective nucleotides.
- This step amplifies a subset of fragments, reducing complexity and improving resolution.
5. Gel Electrophoresis and Analysis
- The amplified fragments are separated on a polyacrylamide gel or using capillary electrophoresis.
- The banding pattern reflects differences in the genetic sequence.
Mechanism of AFLP
- DNA is digested with restriction enzymes.
- Adapters are ligated to the digested DNA fragments.
- PCR amplification is performed using adapter-specific primers.
- Selective primers ensure that only a subset of fragments are amplified.
- The resulting fragments are visualized and analyzed.
Why AFLP is Effective
High Resolution
- Detects polymorphisms at a single-base level.
Genome-Wide Coverage
- AFLP allows for genome-wide analysis without prior knowledge of the sequence.
High Reproducibility
- The technique is highly reproducible due to the strict PCR and digestion conditions.
No Prior Sequence Information Required
- AFLP can be applied to any organism, including those with unsequenced genomes.
Advantages of AFLP
| Advantages | Description |
|---|---|
| High Sensitivity | Detects even small sequence differences. |
| Genome-Wide Screening | Covers large parts of the genome. |
| Versatility | Can be applied to different species and genetic backgrounds. |
| High Throughput | Allows the analysis of multiple samples simultaneously. |
| Cost-Effective | Lower cost compared to whole-genome sequencing. |
Limitations of AFLP
Complexity of Interpretation
- Complex banding patterns may make interpretation difficult.
Dominant Marker System
- AFLP cannot distinguish between homozygous and heterozygous loci.
Requirement for High-Quality DNA
- Poor-quality DNA may lead to inconsistent results.
Applications of AFLP
1. Population Genetics
- Used to study genetic diversity and evolutionary relationships.
2. Genome Mapping
- AFLP is used to construct high-resolution genetic maps.
3. Marker-Assisted Selection
- Helps in identifying markers linked to desirable traits in plants and animals.
4. Microbial Typing
- AFLP is used for identifying and differentiating bacterial and viral strains.
Comparison of AFLP with Other Molecular Markers
| Marker Type | Technique | Polymorphism Type | Applications |
|---|---|---|---|
| AFLP | Digestion + PCR | Fragment length variation | Population genetics, genome mapping |
| SSR (Microsatellites) | PCR | Repeat number variation | Gene mapping, parentage testing |
| RFLP | Digestion + Hybridization | Restriction site variation | Genetic fingerprinting |
| RAPD | PCR | Randomly amplified products | Genetic diversity studies |
Challenges and Future of AFLP
1. Automation and High Throughput
- New platforms are increasing the efficiency of AFLP analysis.
2. Combination with Next-Generation Sequencing
- Combining AFLP with NGS can enhance resolution and accuracy.
3. Enhanced Data Analysis
- Machine learning and bioinformatics tools are improving AFLP data interpretation.
Conclusion
Amplified Fragment Length Polymorphism (AFLP) is a powerful technique for detecting DNA polymorphisms based on restriction enzyme digestion and PCR amplification. Its high sensitivity and genome-wide coverage make it ideal for population genetics, genome mapping, and evolutionary studies. Despite some challenges, AFLP remains a preferred technique for analyzing complex genomes.
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FAQs
Q1. Why is AFLP more sensitive than RFLP?
AFLP combines restriction digestion with PCR, allowing it to detect smaller variations in DNA.
Q2. What is the difference between AFLP and RAPD?
AFLP uses selective primers and restriction digestion, whereas RAPD relies on random primers.
Q3. What are the limitations of AFLP?
AFLP is a dominant marker and cannot differentiate between homozygous and heterozygous loci.
This article was written with insights from Let’s Talk Academy, a top coaching institute for life sciences and biotechnology competitive exams.
91 Comments
Akshay mahawar
March 17, 2025Done 👍
Mahima Sharma
August 24, 2025AFLP is a technique used to detect DNA polymorphisms by cutting genomic DNA with restriction enzymes and amplifying the fragments using PCR
Arushi
March 17, 2025👍👍
Prami Masih
March 19, 2025Okay sir I
Suman bhakar
March 20, 2025👍
pallavi gautam
March 22, 2025doneee
Kabeer Narwal
March 24, 2025👍
Ujjwal
March 25, 2025Done
Komal Sharma
August 23, 2025AFLP is a technique used to detect DNA polymorphisms by cutting genomic DNA with restriction enzymes and amplifying the fragments using PCR. It allows for the detection of genetic variation at a high resolution without prior knowledge of the genome sequence so option 4 is correct
Karishma
August 24, 2025In AFLP, digestion by restriction endo and followed by two steps pcr
Karishma
August 24, 2025In AFLP, digestion by restriction endonuclease and followed by two steps pcr.
Sheikh Aleen
August 24, 2025Digestion of DNA with restriction enzymes followed by two PCR steps
Neelam Sharma
August 24, 2025Digestion of DNA with restriction enzymes followed by two PCR steps. AFLP involves an initial digestion of genomic DNA using restriction enzymes, followed by selective amplification using two PCR steps.
karishma don
August 24, 2025in AFLP digestion of dna by restriction endonuclease and then linkers added to the ends of fragment by which primers are prepared and further amplification of all dna and in the next pcr cycle specific dna with linkers are taken and specific primers are design which are INTERNAL to linker sequence
Sakshi yadav
August 24, 2025Digestion of DNA with RE and followed the 2 steps of PCR
HIMANI FAUJDAR
August 24, 2025Ans correct answer is (4) ,AFLP is a powerful technique in which used for the detecting DNA polymorphisms based on restriction enzyme digestion and PCR amplification.
Divya rani
August 24, 2025AFLP is the two step PCR technique and in this technique we perform digestion of DNA with restriction endonuclease enzyme then link them with linker DNA sequence for amplification of all DNA.
Mohd juber Ali
August 24, 2025The 4th statement is right
Firstly dsDna digest with the help of RE then add specific linker dna on fragments end then denaturation then taq polymerase amplifie dna (so two step of PCR)
Mitali saini
August 24, 2025Correct Answer: (4) Digestion of DNA with restriction enzymes followed by two PCR steps
The correct statement is (4) Digestion of DNA with restriction enzymes followed by two PCR steps. AFLP involves an initial digestion of genomic DNA using restriction enzymes, followed by selective amplification using two PCR steps.
Divya Mandhania
August 24, 2025The 4th statement is right
Nilofar Khan
August 24, 2025Correct Answer: (4) Digestion of DNA with restriction enzymes followed by two PCR step
Ankita Pareek
August 24, 2025Digestion of dna by restriction enzyme followed by two pcr steps
AKANKSHA RAJPUT
August 24, 2025(4) Digestion of DNA with restriction enzymes followed by two PCR steps
Anurag Giri
August 24, 2025Ans 4
Digestion of DNA with restriction enzymes followed by two PCR steps. AFLP involves an initial digestion of genomic DNA using restriction enzymes, followed by selective amplification using two PCR steps.
MOHIT AKHAND
August 24, 2025Ans 4
Done sir ✅
anjani sharma
August 24, 2025Answer 4
AFLP involves an initial digestion of genomic DNA using restriction enzymes, followed by selective amplification using two PCR steps.
Shobha Sharma
August 24, 2025Digestion of DNA with restriction enzyme followed by two PCR step
Priya dhakad
August 24, 2025Digestion of DNA with restriction enzymes followed by two PCR steps.
Roopal Sharma
August 24, 2025Amplified Fragment Length Polymorphism (AFLP) is a powerful technique for detecting DNA polymorphisms based on restriction enzyme digestion and PCR amplification
Dipti Sharma
August 24, 2025Digestion of DNA with restriction enzymes followed by two PCR steps.
First step involves digestion with restriction enzyme and step involves linker dna ligation for amplification.
Khushi Singh
August 24, 2025Digestion of DNA with restriction enzymes followed by two PCR steps
Priyanka Choudhary
August 24, 2025Digestion of DNA with restriction enzymes followed by two PCR steps is correct answer
Priti khandal
August 24, 20254 is correct answer Digestion of DNA with restriction enzyme followed by two pcr steps
Alec
August 24, 2025correct ans. is digestion of Dna with restriction enzymes followed by two PCR steps.
Bharti Yadav
August 24, 2025Digestion of DNA with restriction enzymes followed by two PCR
Kajal
August 24, 2025Digestion of dna by RE (6 cutter) followed by 2step PCR is correct one ….
Mansukh Kapoor
August 24, 2025The correct answer is option 4th
Digestion of DNA with restriction enzymes followed by two PCR steps is the correct statement
Neha Yadav
August 24, 2025Digestion of DNA with restriction enzyme followed by 2 PCR step
Swati Choudhary
August 24, 2025Digestion of DNA with restriction enzymes followed by two PCR steps is the correct statement.
Anjali
August 25, 2025Digestion of DNA with restriction enzymes followed by two PCR steps. Is correct
Surbhi Rajawat
August 25, 2025In AFLP after the digestion from REN. It includes 2 steps of PCR one where all DNA attatched with linker amplifies and the second where we amplify a specific DNA
Yashika Choudhary
August 25, 2025Aflp two pcr step after restrction enzyme digestion
Komal Pareek
August 25, 2025In AFLP dna is attach with linker then amplification starts
Sonam Saini
August 25, 2025Right answer is 4
Dharmpal Swami
August 25, 2025Aflp after the digestion of dna includes two steps first all dna attached with linker and second is specific dna amplification
shruti sharma
August 25, 2025Correct Answer: (4) Digestion of DNA with restriction enzymes followed by two PCR step
Soniya Shekhawat
August 25, 2025AFLP is the two step PCR in which first digestions on the restriction enzyme including DNA attachment by the linker amplifier and second where we amplifier specific DNA.
Konika Naval
August 25, 2025Option 4
Seema
August 25, 2025Digestion of dna with r.e. followed by 2 step of pcr .
Deepika Sheoran
August 26, 2025Option 4th is correct answer
Digestion of DNA with Restriction enzyme followed by two PCR steps.
Muskan singodiya
August 26, 2025Digestion of dna with restrictions enzyme followed by two step pcr
Shivani
August 26, 2025Digestion of DNA with restriction enzymes followed by two PCR steps.
Payal Gaur
August 26, 2025Option 4 digestion of DNA with restriction enzyme followed by two PCR step. In second PCR specific primer are design (Internal restriction site)
Minal Sethi
August 26, 2025Digestion with restriction endonuclease followed by two set of PCR
Mohini
August 26, 2025Digestion of DNA with restriction enzyme followed by two PCR steps.
Kajal
August 26, 2025Digestion of dna with restriction enzymes followed by two PCR steps
Avni
August 26, 2025Digestion of dna with restriction enzymes followed by two PCR steps
Aakanksha Sharma
August 26, 2025Digestion of dna with restriction enzyme followed by 2 PCR step
Aman Choudhary
August 27, 2025Option D is correct
Digestion of DNA with restriction enzymes followed by two PCR steps.
Aafreen Khan
August 27, 2025Digestion of DNA with restriction enzymes followed by two PCR steps
One is attached linker to all DNA and second is amplify the specific DNA
Varsha Tatla
August 27, 2025Aflp is a two PCR step process after digestion of restriction enzyme
Rakesh Dhaka
August 27, 2025Option 4
Digestion of DNA with restriction enzymes followed by two PCR steps is the correct statement
Surendra Doodi
August 27, 2025AFLP is based on the selective amplification of DNA fragments produced by digestion with restriction enzymes.
Sakshi
August 27, 20254 is correct option
Digestion of DNA by restriction enzyme followed by two PCR steps
Devika
August 28, 2025Option D
Kanica Sunwalka
August 28, 2025done
Asha Gurzzar
August 28, 2025In AFLP digestion of dna is followed by two pcr steps by restriction enzyme
Khushi Vaishnav
August 28, 2025Digestion of DNA with restriction enzymes followed by two PCR steps
Sneha Kumawat
August 28, 2025Aflp is molecular technique that combines restriction enzyme digestion and PcR amplification to identify genetic variation
Santosh Saini
August 29, 2025Digestion of DNA with restriction enzymes followed by two PCR steps
Tanvi Panwar
August 29, 2025Digestion of DNA by restriction enzymes followed by 2 PCR steps.
Pooja Sharma Sharma
August 29, 2025Digestion of DNA with restriction enzymes followed by two PCR steps
Aartii sharma
August 29, 2025Digestion of DNA by restriction enzyme followed by 2 PCR steps
Simran Saini
August 29, 2025Digestion of DNA with restriction enzymes followed by two PCR steps.
Arushi Saini
August 30, 2025Digestion of DNA with rstriction enzyme followed by 2 pcr steps
1= adaptor sequence ligation
2= primer against adaptor sequence
Anisha Beniwal
August 30, 2025Digestion of DNA with restriction enzymes followed by two PCR steps.
Neeraj Sharma
August 30, 2025In AFLP we digest the DNA with restriction enzymes followed by 2 pcr steps
Sakshi Kanwar
August 30, 2025Digestion of DNA with restriction enzymes followed by two PCR steps.
Yogita
August 31, 2025Digestion of DNA with restriction enzyme followed by steps
Preeti
September 1, 2025Digestion of DNA with restriction enzymes is followed by 2 pcr steps
Kirti agarwal
August 31, 2025Digested by restrication enzyme in 2 step pcr
Palak Sharma
August 31, 2025Digestion of dna with r.e. followed by 2 step of pcr .
Shrddha modi
August 31, 2025Option D aflp amplified fragments length polymorphism….
1st pcr . adaptor sequence length
2nd pcr . Primer against adaptor sequence
Saniya Jangid
September 1, 2025Done
Shobha Kanwar
September 1, 2025AFLP is a technique used to detect DNA polymorphisms by cutting genomic DNA with restriction enzymes and amplifying the fragments using PCR
Himanshi
September 1, 2025Digestion of DNA with restriction enzymes followed by two PCR steps.
Priyanka
September 2, 2025Firstly digested with 6 and 4 cutter then two step pcr
Priyanka
September 2, 2025Firstly digested with 6 and 4 cutter then two step pcr in aflp
Anisha jakhar
September 4, 2025(4) Digestion of DNA with restriction enzymes followed by two PCR steps
Muskan Yadav
September 4, 2025Digestion of DNA with restriction enzymes followed by two PCR steps
Kiran Kumawat
September 7, 2025Option 4 is correct because DNA is digested by restrictions enzymes (6 cutter) and is followed by two step PCR, ligation of linker ,amplification and selective amplification