title: 2
organism: human oral metagenome
description: deep dentinal caries
attributes.tag: age
attributes.value: 20
attributes.tag: collection_date
attributes.value: 2012
attributes.tag: description
attributes.value: deep dentinal caries
attributes.tag: env_biome
attributes.value: human-associated habitat
attributes.tag: env_feature
attributes.value: human-associated habitat
attributes.tag: env_material
attributes.value: Dentinal Carie
attributes.tag: geo_loc_name
attributes.value: Brazil:Natal
attributes.tag: host
attributes.value: Homo sapiens
attributes.tag: lat_lon
attributes.value: 5.78 N 35.29 E
attributes.tag: organism
attributes.value: human oral metagenome
attributes.tag: sample_title
attributes.value: 2
attributes.tag: seq_methods
attributes.value: "The 16S rRNA gene V4 variable region PCR primers 515/806 (CAPORASO et al., 2011) with barcode on the forward primer were used in a 30 cycle PCR (5 cycle used on PCR products) using the HotStarTaq Plus Master Mix Kit (Qiagen, USA) under the following conditions: 94°C for 3 minutes, followed by 28 cycles of 94°C for 30 seconds, 53°C for 40 seconds and 72°C for 1 minute, after which a final elongation step at 72°C for 5 minutes was performed. After amplification, PCR products are checked in 2% agarose gel to determine the success of amplification and the relative intensity of bands. Multiple samples are pooled together in equal proportions based on their molecular weight and DNA concentrations. Pooled samples are purified using calibrated Ampure XP beads. Then the pooled and purified PCR product is used to prepare DNA library by following Illumina TruSeq DNA library preparation protocol. Paired-end sequencing was performed at MR DNA (www.mrdnalab.com, Shallowater, TX, USA) on a MiSeq following the manufacturer’s guidelines. The sequences from each end were joined after quality q25 trimming of the ends with MR DNA pipeline. "
attributes.tag: sex
attributes.value: male
attributes.tag: BioSampleModel
attributes.value: MIMARKS.survey
attributes.tag: BioSampleModel
attributes.value: MIGS/MIMS/MIMARKS.human-oral