Experiment information
Accession CRX047158
Organism Grus nigricollis
Title DSB_2
BioProject PRJCA001345
BioSample SAMC065573
Platform Illumina MiSeq
Library
Library name Construction protocol Strategy Source Selection Layout
Extraction of DNA from a 1.0 g portion of each fecal sample was performed using the E. Z. N. A. Soil DNA Kit (Omega Bio-tek, Norcross, GA, USA) following the manufacturer's protocol. Extracted DNA was quantified and evaluated for purity using Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and gel electrophoresis, respectively. The V4-V5 hyper-variable regions of the 16S rRNA genes was amplified using the universal primer sequence 515F 5’-(GTGCCAGCMGCCGCGGTAA)-3’ and 909R 5’-(CCCCGYCAATTCMTTTRAGT)-3’. The PCR conditions were as follows: initial denaturation at 95°C for 3 min; first 5 cycles of 95°C denaturation for 30 s, 45°C annealing for 30 s, and 72°C extension for 30 s, then 20 cycles of 95°C denaturation for 30 s, 55°C annealing for 30 s, and 72°C extension for 30 s and a final extension at 72 °C for 5 min. The PCR reaction mixture (total 30 μl) contained 2 x KAPA HiFi HotStart Ready Mix (Kapa Biosystems, Wilmington, MA, USA) 15 μl, 10 μM of each primer and 20 ng of template DNA. Resulting amplicons were purified using an AMPure XP kit (Beckman Coulter Inc., Brea, CA, USA). Resulting DNA were delivered to Sangon BioTech (Shanghai, China) for library construction using universal Illumina adaptor and index. Sequencing was performed using the Illumina MiSeq system (Illumina, CA, USA) according to the manufacturer’s instructions. AMPLICON GENOMIC PCR FRAGMENT
Processing Planned read length (bp): 400
Release date2019-04-12
Run
Run Accession Release date Run Data File Information
File nameFile size (MB)
CRR053043 2019-04-12 DSB_DSB_2_GTCTGG_39443.last.fastq.gz 3.77
SubmitterWen Wang (007cell@163.com)
OrganizationsQinghai University
Date submitted2019-03-29
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