Caetano-Anollés G
Institute of Agriculture, University of Tennessee, Knoxville 37901-1071.
Plant Mol Biol. 1994 Sep;25(6):1011-26. doi: 10.1007/BF00014674.
Multiple arbitrary amplicon profiling (MAAP) uses one or more oligonucleotide primers (> or = 5 nt) of arbitrary sequence to initiate DNA amplification and generate characteristic fingerprints from anonymous genomes or DNA templates. MAAP markers can be used in general fingerprinting as well as in mapping applications, either directly or as sequence-characterized amplified regions (SCARs). MAAP profiles can be tailored in the number of monomorphic and/or polymorphic products. For example, multiple endonuclease digestion of template DNA or the use of mini-hairpin primers can enhance detection of polymorphic DNA. Comparison of the expected and actual number of amplification products produced with primers differing in length, sequence and GC content from templates of varying complexity reveal severe departures from theoretical formulations with interesting implications in primer-template interaction. Extensive primer-template mismatching can occur when using templates of low complexity or long primers. Primer annealing and extension appears directed by an 8 nt 3'-terminal primer domain, requires sites with perfect homology to the first 5-6 nt fom the 3' terminus, and involves direct physical interaction between amplicon annealing sites.
多重任意扩增子分析(MAAP)使用一个或多个任意序列的寡核苷酸引物(≥5个核苷酸)来启动DNA扩增,并从无名基因组或DNA模板生成特征性指纹图谱。MAAP标记可直接用于一般指纹分析以及作图应用,也可作为序列特征化扩增区域(SCARs)使用。MAAP图谱可以在单态和/或多态产物的数量上进行调整。例如,模板DNA的多次核酸内切酶消化或使用小发夹引物可以增强对多态性DNA的检测。将不同长度、序列和GC含量的引物与不同复杂程度的模板产生的预期和实际扩增产物数量进行比较,结果显示与理论公式存在严重偏差,这对引物-模板相互作用具有有趣的启示。使用低复杂度模板或长引物时,可能会出现广泛的引物-模板错配。引物退火和延伸似乎由一个8个核苷酸的3'末端引物结构域引导,需要与3'末端的前5-6个核苷酸具有完美同源性的位点,并且涉及扩增子退火位点之间的直接物理相互作用。