Kolchinsky A M, Funke R P, Gresshoff P M
Plant Molecular Genetics, University of Tennessee, Knoxville 37901-1071.
Biotechniques. 1993 Mar;14(3):400-3.
PCR-type amplification of genomic sequences using short, single and arbitrarily chosen primers (by DNA amplification fingerprinting [DAF]) was developed in our laboratory. Here we show that it is possible to produce specific fingerprints for subgenomic DNA fragments embedded in agarose, including fractions from conventional and pulse field gels. Fractions of restricted genomic DNA differing in mobility by only 2-3 mm gave one to three different products with different single primers. Yeast chromosome specific markers were easily produced and cloned. We also used DAF to generate specific amplification products of DNA cloned into a yeast artificial chromosome. These products were readily cloned into a plasmid vector.
我们实验室开发了使用短的、单个且任意选择的引物通过DNA扩增指纹技术(DAF)对基因组序列进行PCR型扩增的方法。在此我们表明,有可能为嵌入琼脂糖中的亚基因组DNA片段生成特异性指纹图谱,包括来自常规凝胶和脉冲场凝胶的片段。迁移率仅相差2 - 3毫米的限制性基因组DNA片段,使用不同的单个引物可产生一到三种不同的产物。酵母染色体特异性标记物很容易产生并克隆。我们还使用DAF来生成克隆到酵母人工染色体中的DNA的特异性扩增产物。这些产物很容易克隆到质粒载体中。