title: qiime_experiment_939:441290:31
design: Case/Control comparison between infants with and without current wheezing episodes that live in the rural tropics of Ecuador
library_name: A51:31
library_construction_protocol: Quadruplicated PCR products were pooled in 1 per sample, then were cleaned using AMPure XP bead kit and quantified using the Invitrogen Quant iT kit. Number of molecules were calculated per sample according to the Roche Junior 454 titanium Lib-L preparation manual, and the same amount of DNA molecules were multiplexed prior pyrosequencing. A ratio of 0.3 molecules of DNA per bead was used for the emPCR.
attributes.tag: experiment_center
attributes.value: NHLI_IMPERIAL_COLLEGE_LONDON
attributes.tag: run_center
attributes.value: MOLECULAR GENETICS GROUP - NHLI
attributes.tag: chimera_check
attributes.value: chimera slayer
attributes.tag: target_gene
attributes.value: 16S rRNA
attributes.tag: sequencing_meth
attributes.value: pyrosequencing
attributes.tag: run_date
attributes.value: 10/28/10
attributes.tag: pcr_primers
attributes.value: FWD:CCTACGGGAGGCAGCAG;REV:CCGTCAATTCMTTTRAGT
attributes.tag: platform
attributes.value: FASTA
attributes.tag: library_construction_protocol
attributes.value: Quadruplicated PCR products were pooled in 1 per sample, then were cleaned using AMPure XP bead kit and quantified using the Invitrogen Quant iT kit. Number of molecules were calculated per sample according to the Roche Junior 454 titanium Lib-L preparation manual, and the same amount of DNA molecules were multiplexed prior pyrosequencing. A ratio of 0.3 molecules of DNA per bead was used for the emPCR.
attributes.tag: experiment_design_description
attributes.value: Case/Control comparison between infants with and without current wheezing episodes that live in the rural tropics of Ecuador
attributes.tag: study_center
attributes.value: NHLI_IMPERIAL_COLLEGE_LONDON
attributes.tag: library_selection
attributes.value: PCR
attributes.tag: row_number
attributes.value: 31
attributes.tag: sample_center
attributes.value: FEPIS_QUININDE
attributes.tag: pcr_cond
attributes.value: initial denaturation:94degC_2min; 30cycles of: denaturation=94degC_20sec; annealing=50degC_30sec; amplification=72degC_5min; final hold=4degC
attributes.tag: nucl_acid_amp
attributes.value: Polymerase chain reaction (PCR) was used to amplify the variable regions 3 to 5 (V3-V5, Primers 454B_357F and 454A_926R 22) of the gene that encodes for 16S rRNA in bacteria.  Samples were multiplexed using sample specific barcodes23 and Roche 454 adaptor sequences (Roche Diagnostics, Oakland). To minimize PCR nucleotide insertion mistakes, a high fidelity Taq polymerase was used, and samples were amplified in quadruplicate reactions with 35 cycles each and then pooled.
attributes.tag: key_seq
attributes.value: TCAG
attributes.tag: experiment_title
attributes.value: Upper Airways Microbiota in Wheezing and Healthy Infants from the Tropics of Rural Ecuador
attributes.tag: target_subfragment
attributes.value: V35
attributes.tag: region
attributes.value: 1
attributes.tag: multiplex_ident
attributes.value: MID63
attributes.tag: nucl_acid_ext
attributes.value: PMID: 20052448