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based on the AFLP for the integration of physical and genetic mapping procedure
based on the AFLP for the integration of physical and genetic mapping procedure
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机译：基于AFLP的物理和遗传作图程序集成
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摘要
A method for providing a genetic and physical map composed of a genome or part thereof, the method comprising the steps of: (a) providing at least two individual genetic markers for genome or part thereof, preferably in the form of genetic map; (b) characterizing each of the genetic markers by at least a fragment of AFLP identificadomediante obtaining genetic AFLP fingerprints; (c) providing a library of clones comprising fragments genome or part thereof, preferentementeuna library chromosomes artificial such as a BAC or YAC, (d) generating a plurality of mixtures, each containing mixture a plurality of individual clones lagenoteca; (e) generating a DNA fingerprint AFLP for each of the mixtures; (f) identifying the plurality of mixtures, the (s) mixture (s) in which a fragment AFLPidentificado in (b) in the genetic fingerprint of the mixture is present; (g) generating a DNA fingerprinting of AFLP for each of the individual clones (s) mixture (s) identified (s) in (f), and identifying the clone comprising the AFLP fragment identified in (b) in their genetic footprint AFLP (h) generate a contig comprising individual clones identified in step (g) based on data laobtención genetic fingerprinting; (i) repeating steps (f) to (h) for at least a second fragment of AFLP identified in (b) so that elsegundo AFLP fragment or further fragments AFLP marker characterizing a second genetic ounos additional genetic markers; and (j) linking at least two of the contigs obtained in (h), thereby obtaining one physical map and genéticointegrado the part of the genome, or comprising at least two genetic markers, wherein the primer pairs AFLP used in steps (b) and (e) exhibit superior selectivity and lospares AFLP primer pairs in step (g) have a lower selectivity, each primer deAFLP comprising a constant sequence and variable sequence at the 3 'end of sequence, constant dichasecuencia comprising a nucleotide sequence hybridizable with at least a portion of adaptadoradyacente the 3 'end of the restriction fragment to be amplified and further comprises a denucleótidos sequence hybridizable with the remains of the restriction site, comprising said selective variablenucleótidos sequence, said complementary to nucleotides of the 3'-selective nucleotide losfr agmentos of adapter-ligated restriction that are adjacent to the constant sequence in quelos AFLP primers of higher selectivity comprise at least one more than loscebadores selective nucleotide AFLP lower selectivity, and wherein AFLP primers used for elmarcador genetic characterization in step (g) differ from AFLP primers used for genetic fingerprinting delas mixtures in steps (b) and (e) only in the number of selective nucleotides.
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