2.4. Sequence analysis
Pyrotag sequencing reads were separated by their barcodes and extracted from the data set. Sequencing reads containing two or more ambiguous nucleotides, low quality score (average
score b 25), or reads shorter than 300 bp were discarded. The Mothur software package was used to analyze microbial communities. CD-HIT-OTU was used to remove chimeric sequences. Sequences were assigned to groups based on their bar codes, and similar sequences were assigned into
operational taxonomic units (OTUs) at a pairwise identify of 97% using UCLUST. The most abundant
sequence in each OTU, designated as the representative sequence,was classified taxonomically using Ribosomal Database Project (RDP) classifier 2.0.1 (Cole et al., 2009). The Silva rRNA database was used to identify regions of local similarity between sequences.
2.4. Sequence analysisPyrotag sequencing reads were separated by their barcodes and extracted from the data set. Sequencing reads containing two or more ambiguous nucleotides, low quality score (averagescore b 25), or reads shorter than 300 bp were discarded. The Mothur software package was used to analyze microbial communities. CD-HIT-OTU was used to remove chimeric sequences. Sequences were assigned to groups based on their bar codes, and similar sequences were assigned intooperational taxonomic units (OTUs) at a pairwise identify of 97% using UCLUST. The most abundantsequence in each OTU, designated as the representative sequence,was classified taxonomically using Ribosomal Database Project (RDP) classifier 2.0.1 (Cole et al., 2009). The Silva rRNA database was used to identify regions of local similarity between sequences.
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