Other refers to genera each representing Gemcitabine solubility dmso <0.1% of all sequences. BIIB057 research buy sequences not aligning to prokaryotic or human genomes with a ≤ 2 bp mismatch were re-aligned to the human genome with decreased stringency (≤10 bp

mismatch), leaving 32,991,450 sequences for contig assembly (Table  1). Using Ray v1.7 [22], 56,712 contigs were assembled and submitted to the MG-RAST pipeline [21]. Post quality control, 53,785 sequences (94.8%), with a mean length of 160 ± 55 bp, were used for further analysis (Table  1). When the contigs were analyzed using a best hit approach through MG-RAST, they aligned predominantly to the phyla of Proteobacteria (65.1%) and Firmicutes (34.6%, Figure  2). The contigs aligned to 194 known genomes at the genus level, predominantly Pseudomonas (61.1%), Staphylococcus (33.4%) and Streptococcus (0.5%), with the highest level of diversity at the genus level within the Proteobacteria phylum (125 different genera, Figure  2). These results are similar to the best hit analysis performed with the non-assembled sequences in that the majority of sequences

are from Staphylococcus and Pseudomonas, but differ in their proportion (Figure  1). Contigs matching viral genomes were observed (< 0.04%), including phages derived from Pseudomonas and Staphylococcus (Figure  2). Contigs also aligned to the genomes of humans, gorillas, chimps and orangutans, KU55933 research buy likely due to the 60% identity criteria used (Figure  2). The observation of some of the genera, including Staphylococcus, Pseudomonas and Pantoea, was further validated through the presence of their rRNA ORFs (Additional file 3). Table 1 Contig assembly and open reading frame (ORF) prediction of Illumina reads (51 bp) from human milk Sequenced reads (51 bp)

261, 532, 204 Matching human 186,010,988 Matching prokaryotic 1,331,996 Used in contig assembly1 32,991,450 Contigs 56, 712 Post quality control 53,785 Average length (bp) 160 ± 55 Total length (bp) 8,630,997 Predicted ORFs 41, 352 Annotated 33,793 rRNAs 103 Functional category 30,128 Unrecognized Vildagliptin 7,559 1 all sequences not matching the human genome (≤10 bp mismatch). Figure 2 Best hit analysis of open reading frames within human milk. Assembled contigs (56,712) were submitted to MG-RAST for analysis. Contigs aligned to 194 known genomes at the genus level (maximum e-value of 1×10-5, minimum identity of 60%, and minimum alignment length of 45 bp). Color denotes phylum and red bars indicate the number of positive alignments. Open reading frames within human milk A total of 41,352 ORFs were predicted using MG-RAST, of which 82% were annotated (33,793 ORFs), and 18% were unrecognized (7,559 sequences, Table  1). A total of 30,128 ORFs corresponded to a functional category (Figure  3). For example, many ORFs encoded proteins for basic cellular function, including those for respiration (4.2%), cell signaling (4.8%), RNA (7.0%), DNA (2.6%), and amino acid metabolism (5.