Experiment information
Accession CRX032459
Organism Pyrrhocorax pyrrhocorax
BioProject PRJCA001082
BioSample SAMC047580
Platform Illumina MiSeq
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 Accession Release date Run Data File Information
File nameFile size (MB)
CRR036630 2018-11-15 raw.split.3_3.1.fq.gz
SubmitterWen Wang (007cell@163.com)
OrganizationsQinghai University
Date submitted2018-11-12
Related experiments