AFLP - A NEW TECHNIQUE FOR DNA-FINGERPRINTING
AFLP - A NEW TECHNIQUE FOR DNA-FINGERPRINTING
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DOI：
10.1093/nar/23.21.4407
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发表时间：
1995-11-11
影响因子：
14.9
通讯作者：
ZABEAU, M
中科院分区：
文献类型：
--
作者：
VOS, P;HOGERS, R;ZABEAU, M
描述了一种新的DNA指纹技术，称为AFLP。AFLP技术是基于从基因组DNA的总消化物中选择性PCR扩增限制性片段。该技术包括三个步骤：（i）DNA的限制和寡核苷酸接头的连接，（ii）限制片段组的选择性扩增，和（iii）扩增片段的凝胶分析，限制性片段的PCR扩增通过使用衔接子和限制性位点序列作为引物退火的靶位点来实现。选择性扩增通过使用延伸到限制性片段中的引物来实现，仅扩增其中引物延伸与限制性位点侧翼的核苷酸匹配的那些片段。使用这种方法，限制性片段组可以通过PCR可视化，而无需了解核苷酸序列。该方法允许大量限制性片段的特异性共扩增。然而，可以同时分析的片段的数量取决于检测系统的分辨率。通常在变性聚丙烯酰胺凝胶上扩增和检测50-100个限制性片段。AFLP技术为任何来源或复杂性的DNA提供了一种新颖且非常强大的DNA指纹技术。
A novel DNA fingerprinting technique called AFLP is described, The AFLP technique is based on the selective PCR amplification of restriction fragments from a total digest of genomic DNA, The technique involves three steps: (i) restriction of the DNA and ligation of oligonucleotide adapters, (ii) selective amplification of sets of restriction fragments, and (iii) gel analysis of the amplified fragments, PCR amplification of restriction fragments is achieved by using the adapter and restriction site sequence as target sites for primer annealing. The selective amplification is achieved by the use of primers that extend into the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotides flanking the restriction sites, Using this method, sets of restriction fragments may be visualized by PCR without knowledge of nucleotide sequence, The method allows the specific co-amplification of high numbers of restriction fragments. The number of fragments that can be analyzed simultaneously, however, is dependent on the resolution of the detection system, Typically 50-100 restriction fragments are amplified and detected on denaturing polyacrylamide gels, The AFLP technique provides a novel and very powerful DNA fingerprinting technique for DNAs of any origin or complexity.