Amplified fragment length polymorphism 2203789 225225291 2008-07-12T15:42:28Z 70.53.128.50 /* Applications */ link inserted for RAPD {{nofootnotes}} [[Image:Electropherogram trace.jpg|thumb|300px|Example of AFLP Data from a Capillary Electrophoresis Instrument]] '''Amplified fragment length polymorphism PCR''' (or '''AFLP-PCR''' or just '''AFLP''') is a [[polymerase chain reaction|PCR-based]] tool used in [[genetics]] research, [[DNA fingerprint]]ing, and in the practice of [[genetic engineering]]. Developed in the early 1990’s by [http://www.keygene.com '''Keygene'''], AFLP uses [[restriction enzymes]] to cut [[Genome|genomic DNA]], followed by [[ligation]] of complementary double stranded adaptors to the ends of the [[restriction fragment]]s. A subset of the restriction fragments are then amplified using two primers complementary to the adaptor and restriction site fragments. The fragments are visualized on denaturing [[polyacrylamide]] [[electrophoresis|gels]] either through [[autoradiography]] or [[fluorescence]] methodologies. AFLP-PCR is a highly sensitive method for detecting [[Polymorphism (biology)|polymorphism]]s in [[DNA]]. The technique was originally described by Vos and Zabeau in 1993<ref>Vos et al 1993</ref>. The procedure of this technique is divided into three steps: [http://www.keygene.com/keygene/techs-apps/FLASH3.php] #Digestion of total cellular DNA with one or more [[restriction enzyme]]s and ligation of restriction half-site specific adaptors to all restriction fragments. #Selective amplification of some of these fragments with two PCR primers that have corresponding adaptor and restriction site specific sequences. #Electrophoretic separation of amplicons on a gel matrix, followed by visualisation of the band pattern. A variation on AFLP is cDNA-AFLP, which is used to quantify differences in gene expression levels. [http://www.keygene.com/keygene/techs-apps/technologies_gene.php] Another variation on AFLP is TE Display, used to detect transposable element mobility. ==Applications== [[Image:AFLP Clustering Analysis.jpg|thumb|300px|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. There are many advantages to AFLP when compared to other marker technologies including randomly amplified polymorphic DNA ([[RAPD]]), [[restriction fragment length polymorphism]] (RFLP), and [[microsatellite]]s. AFLP not only has higher reproducibility, resolution, and sensitivity at the whole genome level compared to 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 (Meudth & Clarke 2007)<ref>(Meudth & Clarke 2007)</ref>. 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== * [http://www.keygene.com/keygene/techs-apps/index.php AFLP Technology note at Keygene] * [http://www.softgenetics.com/AFLPApplicationNote.pdf AFLP Applications] ==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 [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=7501463] *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. *O’Shea, B., Khare, S., Bliss, K., Klein, P., Ficht, T. A., Adams, L. G., Rice-Ficht, A. C. Genotyping and Characterization of Mycobacterium avium subsp. paratuberculosis Using Amplified Fragment Length Polymorphism. 2004. Journal of Clinical Microbiology. 42:3600-3606 *Meudth & Clarke 2007. Almost Forgotten or Latest Practice? AFLP applications, analyses and advances. Trends in Plant Science Volume 12, Issue 3, March 2007, Pages 106-117. doi:10.1016/j.tplants.2007.02.001 [http://dx.doi.org/10.1016/j.tplants.2007.02.001] ==External links== Software for analyzing AFLP data *[http://www.keygene.com/keygene-products/software/quantarsuite.php KeyGene Quantar Suite] ''Versatile marker scoring software'' *[http://www.softgenetics.com/ SoftGenetics] '''''GeneMarker''''' fragment analysis software Online programs for simulation of AFLP-PCR *[http://www.hpa-bioinfotools.org.uk/aflp.html ALFIE] - BProkaryotes or uploaded sequences *In silico AFLP-PCR for [http://insilico.ehu.es/AFLP prokaryotes],some [http://insilico.ehu.es/AFLP/index.php?mo=eukarya eukaryotes] or [http://insilico.ehu.es/user_seqs/ uploaded sequences] [[Category:Molecular biology]] [[Category:DNA]] [[Category:DNA profiling techniques]] [[de:AFLP]] [[fr:Amplified fragment-length polymorphism]] [[sr:AFLP]] [[uk:Поліморфізм довжини ампліфікованих фрагментів]]