Batched analysis of genotypes.

C LeDuc, P Miller, J Lichter, P Parry
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引用次数: 40

Abstract

Polymorphic microsatellite markers are widely used in molecular analyses. The range of allele sizes and the allele frequencies within a population are important characteristics of the marker. Their determination previously has involved genotyping a large number of individuals. We have developed a technique for defining these characteristics by coamplification of many samples in a DNA pool. Groups of 32 and 42 DNA samples were genotyped and results were compared with those from individual genotype determinations. To improve the accuracy in the estimation of allele frequencies, arithmetic removal of stutter bands was carried out and the consistency of each marker was characterized. This approach was also applied to a group of 94 individuals. All of the work has been done using nonradioactive methods. Potential applications of this technique are in population genetics, high throughput genotyping, and loss of heterozygosity studies.

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基因型批量分析。
多态微卫星标记在分子分析中有着广泛的应用。等位基因大小的变化范围和等位基因频率是该标记的重要特征。他们的测定以前涉及到大量个体的基因分型。我们已经开发了一种技术,通过在DNA池中对许多样品进行共扩增来定义这些特征。对32组和42组DNA样本进行基因分型,并与个体基因分型结果进行比较。为了提高等位基因频率估计的准确性,算法去除了口吃频带,并对每个标记的一致性进行了表征。这种方法也适用于一组94人。所有的工作都是用非放射性方法完成的。该技术的潜在应用是群体遗传学、高通量基因分型和杂合性损失研究。
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Rapid and sensitive analysis of mRNA polyadenylation states by PCR. Identification of 3'-terminal exons from yeast artificial chromosomes. Quantitative analysis of specific mRNA transcripts using a competitive PCR assay with electrochemiluminescent detection. Batched analysis of genotypes. A one-step coupled amplification and oligonucleotide ligation procedure for multiplex genetic typing.
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