title: Biological replicate 2
organism: ant metagenome
attributes.tag: isolation_source
attributes.value: Larvae and pupae replicate 2
attributes.tag: host
attributes.value: Solenopsis invicta
attributes.tag: collection_date
attributes.value: 2014-09
attributes.tag: geo_loc_name
attributes.value: Argentina: Formosa
attributes.tag: lat_lon
attributes.value: 25.30 S 58.40 W
attributes.tag: ref_biomaterial
attributes.value: Primary publication
attributes.tag: samp_collect_device
attributes.value: Hand-collected
attributes.tag: samp_mat_process
attributes.value: Total RNA (200 ng) purified from pooled groups of larvae and pupae was used for mRNA purification with the Illumina TruSeq Stranded mRNA Library Preparation Kit (Catalog # RS-122-2101). The low sample protocol was followed according to the manufacturer's instructions. The RNA fragmentation step was omitted according to the protocol in appendix A to maximize library insert length. Rather than fragmenting the RNA, the sealed plate was incubated at 80oC for 2 minutes to elute the primed mRNA from the RNA purification beads. This omission resulted in RNA fragmentation with an average final library size of 467 bp. Library sizes were determined empirically by microfluidic analysis on an Agilent 2100 Bioanalyzer and quantified using the Quant-iT dsDNA kit with broad range standards (ThermoFisher Scientific, Q-33130). Samples were pooled together at equimolar quantities and sequenced twice using the Illumina MiSeq (2x300) cycle kit with version 3 chemistry. Using the Illumina indices, the data were demultiplexed and the runs combined to assign the data to individual samples. All other procedures were followed according to the manufacturer's instructions.
attributes.tag: Instrument
attributes.value: Illumina MiSeq
attributes.tag: collected by
attributes.value: Sanford D. Porter
attributes.tag: identified by
attributes.value: Sanford D. Porter
attributes.tag: BioSampleModel
attributes.value: Metagenome or environmental