Experiment information
Accession CRX032462
Organism Pyrrhocorax pyrrhocorax
Title Large intestine_3
BioProject PRJCA001083
BioSample SAMC047583
Platform Illumina MiSeq
Library
Library name Construction protocol Strategy Source Selection Layout
Microbial DNA was extracted from 12 samples using the E.Z.N.A. ® stool DNA Kit (Omega Bio-tek, Norcross, GA, USA) according to the manufacturer’s protocols. Extracted DNA was quantified and evaluated for purity using NanoDrop 2000 UV-vis spectrophotometer (Thermo Scientific, Wilmington, USA) and 1% agarose gel electrophoresis, respectively. The V4 - V5 hyper-variable region of the 16S rRNA genes was amplified with the universal primers 515F 5′-(GTGCCAGCMGCCGCGG)-3′ and 907R 5′-(CCGTCAATTCMTTTRAGTTT)-3′ by thermocycler PCR system (GeneAmp 9700, ABI, USA). The PCR conditions were as follows: initial denaturation at 95°C for 3 min; 95°C denaturation for 30 s, 55°C annealing for 30 s, and 72°C extension for 45 s, which is repeated for 27 cycles; and a final extension at 72°C for 10 min. The PCR reactions were performed in triplicate and conducted with 20 µl mixture containing 4 µl of 5 X FastPfu buffer, 2 µl of 2.5 mM dNTPs, 0.8 µl of each primer (5 mM), 0.4 µl of FastPfu polymerase and 10 ng of template DNA. The resulted PCR products were extracted from 2% agarose gel and further purified using the AxyPrep DNA Gel Extraction Kit (Axygen Biosciences, Union City, CA, USA) and quantified using QuantiFluor TM - ST (Promega, Madison, Wisconsin, USA) following the manufacturer’s protocols. Purified amplicons were pooled in equimolar and paired-end sequenced (2 X 300 bp) on an Illumina Miseq platform (Illumina, San Diego, USA) according to the standard protocols by Majorbio Bio-Pharm Technology Co. Ltd. (Shanghai, China). CLONE GENOMIC PCR PAIRED
Processing Planned read length (bp) for mate 1: 300
Planned read length (bp) for mate 2: 300
Insert size (bp): 300
Release date2018-11-15
Run
Run Accession Release date Run Data File Information
File nameFile size (MB)
CRR036633 2018-11-15 raw.split.3_4.1.fq.gz
raw.split.3_4.2.fq.gz
3.54
5.45
SubmitterWen Wang (007cell@163.com)
OrganizationsQinghai University
Date submitted2018-11-12
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