Needle's Eye Depth Core (Control) Ion Torrent FASTQ Data (NERC Grant NE/L000326/1)
Soil depth core collected from the Needle’s Eye site in Dumfries, Scotland. Clear plastic depth core was lowered into a bog within the site, excised, and capped at the top and bottom. Core was sliced at 1 cm intervals at the University of Manchester in an anaerobic bag. A total of 42 samples were generated. Soil samples were returned to Newcastle University.
Leech Building, Faculty of Medical Sciences, Framlington Place,
Newcastle,
NE2 4HH
email:
not available
Role: originator
University of Newcastle
Peter Leary
Leech Building, Faculty of Medical Sciences, Framlington Place,
Newcastle,
NE2 4HH
email:
not available
Role: point of contact
University of Newcastle
Dr Neil Gray
Agriculture Building,
Newcastle,
NE1 7RU
email:
not available
Role: principal investigator
Data Quality
DNA was extracted using FastDNA SPIN Kit for Soil. PCR using the 515F and 926R primers targeting the V4-V5 region of the 16S rRNA gene. Amplicon length was 481 bp. PCRs were 25 uL reactions with 22 uL of MegaMix Blue, 1uL of forward and 1 uL of reverse primer at 100 pM concentration, and 1 uL of 1:10 diluted template DNA. Initial denaturation was 95C for 5 minutes, followed by 30 cycles of 95C for 1 minute, 55C for 1 minutes, 72C for 1 minute, and a final elongation step of 72C for 10 minutes. Each depth sample was run in triplicate and pooled. Pooled amplicons were cleaned using Agencourt AMPure XP. PCR amplicons were quantified via Qubit 3.0 fluorometer and a total dilution factor was calculated for each sample, diluting each sample to 1000. Equimolar samples were pooled into a final library. Sequencing was performed on the Ion Torrent PGM on a 316 chip.
INSPIRE Implementing rules laying down technical arrangements for the interoperability and harmonisation of Geology
Commission Regulation (EU) No 1089/2010 of 23 November 2010 implementing Directive 2007/2/EC of the European Parliament and of the Council as regards interoperability of spatial data sets and services
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