Evolution and Biodiversity, Institute of Evolutionary Biology and Environmental Studies, Winterthurerstrasse 190, CH 8057 Zurich, Switzerland.
Mol Ecol Resour. 2011 May;11(3):494-502. doi: 10.1111/j.1755-0998.2010.02957.x. Epub 2010 Dec 28.
The amplified fragment length polymorphism (AFLP) technique is a widely used multi-purpose DNA fingerprinting tool. The ability to size-separate fluorescently labelled AFLP fragments on a capillary electrophoresis instrument has provided a means for high-throughput genome screening, an approach particularly useful in studying the molecular ecology of nonmodel organisms. While the 'per-marker-generated' costs for AFLP are low, fluorescently labelled oligonucleotides remain costly. We present a cost-effective method for fluorescently end-labelling AFLPs that should make this tool more readily accessible for laboratories with limited budgets. Both standard fluorescent AFLPs and the end-labelled alternatives presented here are repeatable and produce similar numbers of fragments when scored using both manual and automated scoring methods. While it is not recommended to combine data using the two approaches, the results of the methods are qualitatively comparable, indicating that AFLP end-labelling is a robust alternative to standard methods of AFLP genotyping. For researchers commencing a new AFLP project, the AFLP end-labelling method outlined here is easily implemented, as it does not require major changes to PCR protocols and can significantly reduce the costs of AFLP studies.
扩增片段长度多态性（AFLP）技术是一种广泛使用的多用途 DNA 指纹图谱工具。在毛细管电泳仪器上对荧光标记的 AFLP 片段进行大小分离的能力为高通量基因组筛选提供了一种方法，这种方法特别适用于研究非模式生物的分子生态学。虽然 AFLP 的“每个标记生成”成本较低，但荧光标记的寡核苷酸仍然昂贵。我们提出了一种经济高效的 AFLP 荧光末端标记方法，这应该使预算有限的实验室更容易获得这种工具。标准荧光 AFLP 和本文提出的末端标记替代物都是可重复的，并且使用手动和自动评分方法评分时会产生相似数量的片段。虽然不建议使用这两种方法组合数据，但这两种方法的结果在质量上是可比的，这表明 AFLP 末端标记是 AFLP 基因分型标准方法的可靠替代方法。对于开始新的 AFLP 项目的研究人员来说，这里概述的 AFLP 末端标记方法易于实施，因为它不需要对 PCR 方案进行重大更改，并且可以显著降低 AFLP 研究的成本。
