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Encyclopedia > Amplified fragment length polymorphism

Amplified fragment length polymorphism PCR, or "AFLP-PCR" (often AFLP), is a tool used in the study of genetics and in the practice of genetic engineering. For a non-technical introduction to the topic, please see Introduction to genetics. ... An iconic image of genetic engineering; this autoluminograph from 1986 of a glowing transgenic tobacco plant bearing the luciferase gene, illustrating the possibilities of genetic engineering. ...

Example of AFLP Data from a Capillary Electrophoresis Instrument
Example of AFLP Data from a Capillary Electrophoresis Instrument

Amplified Fragment Length Polymorphism (AFLP) is a polymerase chain reaction (PCR) based genetic fingerprinting technique that was developed in the early 1990’s by Keygene. AFLP uses restriction enzymes to cut genomic DNA, followed by ligation of complementary double stranded adaptors to the ends of the restriction fragments. A subset of the restriction fragments are then amplified using 2 primers complementary to the adaptor and restriction site fragments. The fragments are visualized on denaturing polyacrylamide gels either through autoradiographic or fluorescence methodologies. Image File history File links No higher resolution available. ... Image File history File links No higher resolution available. ...


AFLP-PCR is a highly sensitive method for detecting polymorphisms in DNA.The technique was originally described by Vos and Zabeau in 1993. The procedure of this technique is divided into three steps: In biology, polymorphism can be defined as the occurrence in the same habitat of two or more forms of a trait in such frequencies that the rarer cannot be maintained by recurrent mutation alone. ... The structure of part of a DNA double helix Deoxyribonucleic acid (DNA) is a nucleic acid that contains the genetic instructions for the development and function of living organisms. ...

  1. Digestion of total cellular DNA with one or more restriction enzymes and ligation of restriction half-site specific adaptors to all restriction fragments.
  2. Selective amplification of some of these fragments with two PCR primers that have corresponding adaptor and restriction site specific sequences.
  3. Electrophoretic separation of amplicons on a gel matrix, followed by visualisation of the band pattern.

A variation on AFLP is TE Display, used to detect transposable element mobility. A restriction enzyme (or restriction endonuclease) is an enzyme that cuts double-stranded DNA. The enzyme makes two incisions, one through each of the sugar-phosphate backbones (i. ...

Contents

Applications

AFLP Phylogeny Analysis Using a Dendrogram
AFLP Phylogeny Analysis Using a Dendrogram

The AFLP technology has the capability to detect various polymorphisms in different genomic regions simultaneously. It is also highly sensitive and reproducible. As a result, AFLP has become widely used for the identification of genetic variation in strains or closely related species of plants, fungi, animals, and bacteria. The AFLP technology has been used in criminal and paternity tests, in population genetics to determine slight differences within populations, and in linkage studies to generate maps for QTL analysis. Image File history File links Size of this preview: 800 × 416 pixelsFull resolution (886 × 461 pixel, file size: 75 KB, MIME type: image/jpeg)AFLP Dendrogram from GeneMarker Software I, the creator of this work, hereby release it into the public domain. ... Image File history File links Size of this preview: 800 × 416 pixelsFull resolution (886 × 461 pixel, file size: 75 KB, MIME type: image/jpeg)AFLP Dendrogram from GeneMarker Software I, the creator of this work, hereby release it into the public domain. ...


There are many advantages to AFLP when compared to other marker technologies including randomly amplified polymorphic DNA (RAPD), restriction fragment-length polymorphism (RFLP), and microsatellites. AFLP not only has higher reproducibility, resolution, and sensitivity at the whole genome level than other techniques, but it also has the capability to amplify between 50 and 100 fragments at one time. In addition, no prior sequence information is needed for amplification. As a result, AFLP has become extremely beneficial in the study of taxa including bacteria, fungi, and plants, where much is still unknown about the genomic makeup of various organisms.


See also

In molecular biology, the term restriction fragment length polymorphism (or RFLP, often pronounced rif-lip) is used in two related contexts: as a characteristic of DNA molecules (arising from their differing nucleotide sequences) by which they may be distinguished, as the laboratory technique which uses this characteristic to compare DNA...

References

  • Zabeau, M and P. Vos. 1993. Selective restriction fragment amplification: a general method for DNA fingerprinting. European Patent Office, publication 0 534 858 A1, bulletin 93/13.
  • Vos, P., Hogers, R., Bleeker, M., et al. 1995. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Research 23(21):4407-4414 [1]
  • Van den Broeck et al., 1998
  • Casa et al., 2000
  • Biedler et al., 2003
  • Ulrich G. Mueller and LaReesa Wolfenbarger. 1999. AFLP Genotyping and fingerprinting.Tree. Vol14. 10:389 - 394.

External links

Software for analyzing AFLP data

  • SoftGenetics GeneMarker® fragment analysis software

Online programs for simulation of AFLP-PCR

  • ALFIE - BProkaryotes or uploaded sequences
  • In silico AFLP-PCR for prokaryotes,some eukaryotes or uploaded sequences

  Results from FactBites:
 
Amplified fragment length polymorphism - Wikipedia, the free encyclopedia (200 words)
Amplified fragment length polymorphism PCR, or "AFLP-PCR" (often AFLP), is a tool used in the study of genetics and in the practice of genetic engineering.
AFLP-PCR is a highly sensitive method for detecting polymorphisms in DNA.The technique was originally described by Zabeau and Vos in 1993.
A variation on AFLP is TE Display, used to detect transposable element mobility.
Restriction fragment length polymorphism - Wikipedia, the free encyclopedia (239 words)
In molecular biology, the term restriction fragment length polymorphism (or RFLP) is used in two related contexts: as a characteristic of DNA molecules (arising from their differing nucleotide sequences) by which they may be distinguished, and as the laboratory technique which uses this characteristic to compare DNA molecules.
The DNA is then cut into restriction fragments by endonucleases, which only cut where there are specific DNA sequences recognized by the enzymes.
The restriction fragments are then separated according to length by agarose gel electrophoresis.
  More results at FactBites »

 
 

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