artificial and natural duplicates in pyrosequencing reads of metagenomic data人工和自然重复在焦磷酸测序读的宏基因组数据.pdfVIP

Niu et al. BMC Bioinformatics 2010, 11:187 /1471-2105/11/187 R E S E A R C H A R T I C L E Open Access Research article Artificial and natural duplicates in pyrosequencing reads of metagenomic data 1 1 2 1,2 Beifang Niu , Limin Fu , Shulei Sun and Weizhong Li* Abstract Background: Artificial duplicates from pyrosequencing reads may lead to incorrect interpretation of the abundance of species and genes in metagenomic studies. Duplicated reads were filtered out in many metagenomic projects. However, since the duplicated reads observed in a pyrosequencing run also include natural (non-artificial) duplicates, simply removing all duplicates may also cause underestimation of abundance associated with natural duplicates. Results: We implemented a method for identification of exact and nearly identical duplicates from pyrosequencing reads. This method performs an all-against-all sequence comparison and clusters the duplicates into groups using an algorithm modified from our previous sequence clustering method cd-hit. This method can process a typical dataset in ~10 minutes; it also provides a consensus sequence for each group of duplicates. We applied this method to the underlying raw reads of 39 genomic projects and 10 metagenomic projects that utilized pyrosequencing technique. We compared the occurrences of the duplicates identified by our method and the natural duplicates made by independent simulations. We observed that the duplicates, including both artificial and natural duplicates, make up 4- 44% of reads. The number of natural duplicates highly correlates with the samples read density (number of reads divided by genome size). For high-complexity metagenomic samples lacking dominant species, natural duplicates only make up 1% of all duplicates. But for some