SAMN14484714
External Id
Astrobiology Center Inta
INSDC center name
2020-03-30T00:00:00Z
INSDC first public
2020-03-30T06:44:04.792Z
INSDC last update
live
INSDC status
Metagenome or environmental
NCBI submission model
Metagenome.environmental.1.0
NCBI submission package
2017-09-01
collection date
Illumina sequencing was applied to the nine replicates of glacial ice: MP1, MP10, MP20, MP30, MP40, MP50, MP70, MP80 and MP100. Purified DNA was quantified and 1ng of input DNA was used in a first PCR of 20 cycles with Q5® Hot Start High-Fidelity DNA Polymerase in the presence of 100nM primers for 16S and 18S amplification. After the first PCR, a second PCR of 15 cycles was performed with Q5® Hot Start High-Fidelity DNA Polymerase in the presence of 400nM of primers of the Access Array Barcode Library for Illumina Sequencers. The finally obtained amplicons were validated and quantified by Bioanalyzer and an equimolecular pool was purified using AMPure beads and titrated by quantitative PCR using the Kapa-SYBR FAST qPCR kit for LightCycler 480 and a reference standard for quantification. The pool of amplicons were denatured prior to be seeded on a flowcell at a density of 10pM, where clusters were formed and sequenced using a MiSeq Reagent Kit v3, in a 2x300 pair-end sequencing run on a MiSeq sequencer to obtain 100000 reads per sample approximately. Genomic data were analyzed with Base Space platform.
description
glacier ice
isolation source
42.6753700 N 0.0339000 E
lat lon
glacier metagenome
http://purl.obolibrary.org/obo/NCBITaxon_1651087
organism
Metagenome or environmental sample from glacier metagenome
title