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SRX2530009: "UCE target enrichment of Brachyhypopomus_occidentalis_LSUMZ14836: fin/muscle tissue"
1 ILLUMINA (Illumina MiSeq) run: 608,034 spots, 304.8M bases, 188.5Mb downloads

Design: We input 100-1000 ng sheared DNA to a modified genomic DNA library preparation protocol (v1.10; http://ultraconserved.org) that incorporated "with-bead" cleanup steps (Fisher et al. 2011) using a generic SPRI substitute (hereafter SPRI; Rohland and Reich 2012). Following adapter ligation, 50% of the resulting library volume (~15 _L; 50-500 ng) was PCR amplified using a reaction mix of 25 _L HiFi HotStart polymerase (Kapa Biosystems), 5 _L of Illumina TruSeq primer mix (5 _M each), and 5 _L ddH20 using the following thermal protocol: 98ĄC for 45s; 10-16 cycles of 98ĄC for 15s, 60ĄC for 30s, 72ĄC for 60s; and a final extension of 72ĄC for 5m. Resulting reactions were purified using 1X SPRI, and we re-hydrated libraries in 33 _L ddH2O. We quantified 2 _L of each library using a Qubit fluorometer. Groups of eight libraries at equimolar ratios were combined into enrichment pools having a final concentration of 147 ng/_L. Using a custom target enrichment bait kit from MYcroarray Inc. (Faircloth et al. 2012; McGee et al. 2016) a standard UCE enrichment protocol was followed (v1.4; http://ultraconserved.org). Based on the size-adjusted concentrations estimated by qPCR, we created an equimolar pool of libraries at 10 nM concentration, and we sequenced 9-10 pmol of each pool-of-pooled libraries using either (Supplemental Table 2) paired-end, 250 bp sequencing on an Illumina MiSeq (v2; UCLA Genotyping Core Facility) or paired-end 100 bp sequencing on an Illumina HiSeq (v3; UCLA Neuroscience Genomics Core).
Submitted by: Louisiana State University
Study: Phylogenomic Systematics of Ostariophysan fishes: Ultraconserved Elements Support the Surprising Non-monophyly of Characiformes
show Abstracthide Abstract
Ostariophysi is a superorder of bony fishes including more than 10,200 species in 1,100 genera and 70 families. This superorder is traditionally divided into five major groups (orders): Gonorynchiformes (milkfishes and sandfishes), Cypriniformes (carps and minnows), Characiformes (tetras and their allies), Siluriformes (catfishes), and Gymnotiformes (electric knifefishes). Unambiguous resolution of the relationships among these lineages remains elusive, with previous molecular and morphological analyses failing to produce a consensus phylogeny. In this study, we use over 350 ultraconserved element (UCEs) loci comprising five million base pairs collected across thirty-five representative ostariophysan species to compile one of the most data-rich phylogenies of fishes to date. We use these data to infer higher-level (interordinal) relationships among ostariophysan fishes, focusing on the monophyly of the Characiformes— one the most contentiously debated groups in fish systematics. As with most previous molecular studies, we recover a non-monophyletic Characiformes with the two monophyletic suborders, Citharinoidei and Characoidei, more closely related to other ostariophysan clades than to each other. We also explore incongruence between results from different UCE datasets, issues of orthology, and the use of morphological characters in combination with our molecular data.
Sample:
SAMN06273915 • SRS1950909 • All experiments • All runs
Library:
Name: Brachyhypopomus_occidentalis_LSUMZ14836
Instrument: Illumina MiSeq
Strategy: WGS
Source: GENOMIC
Selection: Hybrid Selection
Layout: PAIRED
Runs: 1 run, 608,034 spots, 304.8M bases, 188.5Mb
Run# of Spots# of BasesSizePublished
SRR5220070608,034304.8M188.5Mb2017-02-05

ID:
3661164

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