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Author:
Date:
2015-05-20
[Abstract] Single Nucleotide Polymorphisms (SNPs), which constitute single base-pair variations in the DNA sequence, are the most abundant molecular markers in plant and animal genomes. They are becoming the markers of choice for genotyping in all fields of molecular biology, as they are easily prone to automation and high throughput, for example through pyrosequencing. This technology is accurate, flexible and can be easily automated. However, the need for primers labelled with biotin, promptly rise the cost of any methodology employing a pyrosequencing approach. In this protocol we described an improved, efficient, reliable and cost-effective pyrosequencing protocol, based on a universal M13 biotinylated primer, for SNP genotyping in plants.
[摘要] 单核苷酸多态性（SNP），其构成DNA序列中的单碱基对变异，是植物和动物基因组中最丰富的分子标记。 它们正成为分子生物学所有领域中基因分型的选择标记，因为它们易于自动化和高通量，例如通过焦磷酸测序。 这种技术是准确，灵活和可以很容易自动化。 然而，对用生物素标记的引物的需要迅速提高了使用焦磷酸测序方法的任何方法的成本。 在这个协议中，我们描述了基于通用M13生物素化引物，用于植物中的SNP基因分型的改进的，有效的，可靠的和成本有效的焦磷酸测序方案。
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