SAMN54248049
External Id
Astrobiology Center Inta
INSDC center name
2025-12-22T00:00:00Z
INSDC first public
2025-12-22T04:59:22.330Z
INSDC last update
SRS27541330
INSDC secondary accession
live
INSDC status
Metagenome or environmental
NCBI submission model
Metagenome.environmental.1.0
NCBI submission package
SRS27541330
SRA accession
2022-09
collection_date
Illumina sequencing was performed on five ice samples: SOM1, SOM2, SOM3, SOM4, and SOM5. Purified DNA was quantified, and 1 ng of input DNA was used for an initial PCR of 20 cycles with Q5 Hot Start High-Fidelity DNA Polymerase and 100 nM primers targeting the 16S region. A second PCR of 15 cycles was then carried out using the same polymerase and 400 nM primers from the Access Array Barcode Library for Illumina Sequencers. The resulting amplicons were validated and quantified with a Bioanalyzer, and an equimolar pool was purified using AMPure beads. This pool was titrated by quantitative PCR with the KAPA SYBR FAST qPCR kit for LightCycler 480, using a reference standard for accurate quantification. Amplicons were denatured and loaded onto a flow cell at a concentration of 10 pM, where cluster generation and sequencing were performed with a MiSeq Reagent Kit v3 in a 2 300 bp paired-end run on a MiSeq system, yielding approximately 100,000 reads per sample. Genomic data were analyzed using the BaseSpace platform.
description
Spain: Huesca
geo_loc_name
Ice cave
isolation_source
42.733055 N 0.190427 W
lat_lon
ice metagenome
http://purl.obolibrary.org/obo/NCBITaxon_452919
organism
SOM_2__16S
title