Genome sequencing information Genome project history The organism was selected for sequencing on the basis of its phylogenetic position selleck and 16S rRNA similarity to other members of the genus Clostridium, and is part of a ��culturomics�� study of the human digestive flora aiming at isolating all bacterial species within human feces. It was the 74th genome of a Clostridium species and the first genome of Clostridium senegalense sp. nov. The Genbank accession number is “type”:”entrez-nucleotide”,”attrs”:”text”:”CAEV00000000″,”term_id”:”379048610″,”term_text”:”CAEV00000000″CAEV00000000 and consists of 191 contigs. Table 2 shows the project information and its association with MIGS version 2.0 compliance. Table 2 Project information Growth conditions and DNA isolation C. senegalense sp. nov.

strain JC122T, CSUR P152 = DSM 25507, was grown on blood agar medium at 37��C. Five petri dishes were spread and resuspended in 5×100��l of G2 buffer (EZ1 DNA Tissue kit, Qiagen). A first mechanical lysis was performed by glass powder on the Fastprep-24 device (Sample Preparation system) from MP Biomedicals, USA) using 2×20 seconds cycles. DNA was then treated with 2.5 ��g/��L lysozyme (30 minutes at 37��C) and extracted through the BioRobot EZ 1 Advanced XL (Qiagen). The DNA was then concentrated and purified on a Qiamp kit (Qiagen). The yield and the concentration was measured by the Quant-it Picogreen kit (Invitrogen) on a Genios_Tecan fluorometer at 70.7 ng/��l. Genome sequencing and assembly This project was loaded twice on a 1/4 region for the paired end application and once on a 1/8 region for the shotgun on PTP Picotiterplates.

The shotgun library was constructed with 500ng of DNA as described by the manufacturer Roche with the GS Rapid library Prep kit. For the paired-end sequencing, DNA (5��g) was mechanically fragmented on the Hydroshear device (Digilab, Holliston, MA, USA) with an enrichment size of 3-4kb. The DNA fragmentation was visualized using an Agilent 2100 BioAnalyzer on a DNA labchip 7500 to yield an optimal size of 3.6 kb. The library was constructed according to the 454_Titanium paired end protocol and manufacturer. Circularization and nebulization were performed and generated a pattern with an optimum at 561 bp. After PCR amplification through 15 cycles followed by double size selection, the single stranded paired end library was then quantified on the Quant-it Ribogreen kit (Invitrogen) on the Genios_Tecan fluorometer at 52pg/��L.

The library concentration equivalence was calculated as 1.7E+08 molecules/��L. The library was held at -20��C until use. The shotgun library was clonally amplified Dacomitinib with 3cpb in 3 emPCR reactions and the paired end library was amplified with lower cpb (1cpb) in 4 emPCR reactions with the GS Titanium SV emPCR Kit (Lib-L) v2. The yield of the emPCR was 5.37% for the shotgun and 19.