Comparative analysis of human saliva microbiome diversity by barcoded pyrosequencing and cloning approaches
Comparative analysis of human saliva microbiome diversity by barcoded pyrosequencing and cloning approaches
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DOI：
10.1016/j.ab.2009.04.034
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发表时间：
2009-08-01
影响因子：
2.9
通讯作者：
Stoneking, Mark
中科院分区：
文献类型：
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作者：
Nasidze, Ivan;Quinque, Dominique;Stoneking, Mark
宏基因组研究传统上依赖于克隆聚合酶链反应（PCR）产物和测序多个克隆。然而，这种方法是繁琐和昂贵的，从而限制了可以解决的问题的范围和规模。DNA测序技术的最新发展使得能够通过以有限的样品处理和较低的成本对许多样品进行并行深入分析来显著增加通量。我们直接将传统的克隆方法与条形码焦磷酸测序方法进行了比较，以确定后者是否准确地描述了人类唾液中的微生物组多样性。我们的结果表明，尽管读长较短，焦磷酸测序方法提供了与基于传统克隆和测序方法的结果非常一致的人类唾液微生物组的描述。(C)2009 Elsevier Inc. All rights reserved.
Metagenomic studies traditionally rely on cloning polymerase chain reaction (PCR) products and sequencing multiple clones. However, this approach is tedious and expensive, thereby limiting the range and scale of questions that can be addressed. Recent developments in DNA sequencing technologies enable a dramatic increase in throughput via parallel in-depth analysis of many samples with limited sample processing and lower costs. We directly compared the traditional cloning approach with a barcoded pyrosequencing method to see whether the latter accurately describes microbiome diversity in human saliva. Our results indicate that despite the shorter read lengths, the pyrosequencing approach Provides a description of the human salivary microbiome that is in good agreement with results based on the traditional cloning and sequencing approach. (C) 2009 Elsevier Inc. All rights reserved.