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SRX1745355: 16S rRNA sequencing of all variable regions for a low complexity, marine biocathode biofilm, Biocathode MCL
1 ILLUMINA (Illumina MiSeq) run: 670,565 spots, 201.4M bases, 143.9Mb downloads

Design: The V3 regions of the 16S rRNA gene were amplified using a two-step PCR amplicon approach recommended by Illumina. Two adapters (forward 5’-TCG TCG GCA GCG TCA GAT GTG TAT AAG AGA CAG, reverse 3’-GTC TCG TGG GCT CGG AGA TGT GTA TAA GAG ACA G) were added to previously published primers (Bartram, Lynch et al. 2011, Cai, Ye et al. 2013). The first PCR reactions were carried out as previously reported in a 25 mL reaction volume using 1 ng genomic DNA as template. The second PCR with index primers was carried out using 10 mL of first PCR products as template with 5 mL each of Illumina index primers in a 50 mL reaction volume. PCR was performed with initial incubation at 72oC for 3 minutes, followed by 10 cycles of 98oC, 10”, 63oC, 30”, 72oC, 30”. Microbial 16S rDNA gene sequences of the variable regions of the 16S rDNA gene were acquired on a MiSEQTM instrument under automated software control (version 2.2.0, Illumina, San Diego, CA).
Submitted by: US Naval Research laboratory
Study: Microbial consortium enriched at the cathode of a solar microbial fuel cell
show Abstracthide Abstract
The objective of this project is to study a microbial biocathode consortium enriched at the U.S. Naval Research Laboratory's Center for Bio/Molecular Science & Engineering. The initial inoculum for the solar microbial fuel cell was seawater collected near Rutgers, NJ, and the community has subsequently been maintained electroautotrophically via serial transfers on cathodes poised at 310 mV vs. SHE. This microbial consortium contains autotrophic organisms which can use a cathode as an electron donor and O2 as an electron acceptor, and has electrochemical properties potentially useful in bioenergy applications including microbial electrosynthesis and microbial fuel cell technology. System biology approaches will be used to identify functional genes that are relevant for bioenergy production, such as fixing CO2 as sole carbon source, electrosynthesis, O2 tolerance, etc. Several heterotrophic constituents have been isolated in pure culture, and have been sequenced by either Illumina or PacBio technologies. Furthermore, closed genomes and methylation data were obtained from PacBio sequencing of metagenomic DNA. Metagenomic libraries generated from eight biological replicates of an enriched biocathode microbial community were sequenced at 2x100 base pairs (bp) (paired-end reads) using an Illumina HiSeq 2000. The reads were assembled using either IDBAUD or Ray. The ideal k-mer length and node coverage were selected based on which gave results most similar to de novo sequencing of isolated bacterial cultures from the enriched community (Marinobacter sp. strain CP1 and Labrenzia sp. strain CP4). Metatranscriptomic libraries were generated from four reactors inoculated with the same inoculum were grown at a set potential of 310 mV SHE until current density stabilized, cyclic voltammetry was recorded, and then two of them were set at a more positive potential (470 mV) and two remained at 310 mV for 48 hours before samples were harvested for RNA extraction. This experiment was repeated with four more reactors inoculated with a new cell suspension from the same source electrode for a total of four biological replicates at each potential.
Sample: Metagenome samples from biocathode biofilm microbial consortium
SAMN04934627 • SRS1424240 • All experiments • All runs
Library:
Name: V3-2
Instrument: Illumina MiSeq
Strategy: AMPLICON
Source: METAGENOMIC
Selection: PCR
Layout: PAIRED
Runs: 1 run, 670,565 spots, 201.4M bases, 143.9Mb
Run# of Spots# of BasesSizePublished
SRR3479899670,565201.4M143.9Mb2017-11-30

ID:
2503005

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