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SRX20108312: UCE capture of frog genomic DNA
1 ILLUMINA (Illumina HiSeq 4000) run: 8.5M spots, 2.4G bases, 989.3Mb downloads

Design: We extracted DNA from tissue samples using the Qiagen DNEasy Blood & Tissue Kit. DNA concentration was quantified using a Qubit DNA BR assay (Life Technologies), and 1500 ng of DNA per sample was sheared using a BioRuptor (Diagenode). Initially all samples were subjected to 10 cycles (30 sec on/30 sec off) to achieve a target size of 400600 bp. Fragmentation was assessed using a BioAnalyzer 2100 (Agilent). Several samples required an additional 8 or 16 cycles for fragmentation. We prepared libraries from the fragmented DNA using the Kapa HyperPrep kit (Roche). We followed the standard protocol, but used Serapure beads for cleanup steps and TruSeq DNA unique dual index adapters (IDT for Illumina). We used the myBaits UCE Tetrapods 5Kv1 probe set (8-reaction kit size, Arbor Biosciences) for captures, following the v4.01 protocol. We pooled 67 libraries for each capture reaction (1500 ng total input DNA) and performed hybridization for 24 hrs. Post-capture PCR amplification used 12 cycles, and the post-cleanup product was eluted in 12 L of buffer EB (10mM Tris-HCl). The eight capture products were pooled in equimolar amounts and sequenced on one lane of an Illumina HiSeq4000 with 150 bp paired-end (PE) reads.
Submitted by: California Academy of Sciences
Study: UCE capture experiment for ranoid and archaeobatrachian frogs
show Abstracthide Abstract
The data available for reconstructing molecular phylogenies has become wildly disparate. Phylogenomic studies can have data for thousands of loci for dozens of species, but for hundreds of other taxa, data may be from only a few genes. Is it possible to integrate these two types of data to combine the advantages of both, addressing the relationships of hundreds of species with thousands of genes? Here we show that this is possible, using data from frogs. We generated a UCE phylogenomic dataset for 138 ingroup species and 3,784 loci, including new data from 49 species. We also assembled a supermatrix dataset, including data from almost every frog genus, with 1-307 genes per taxon. We then produced a combined phylogenomic-supermatrix dataset (gigamatrix) containing 441 ingroup taxa and 4,091 genes, but with 86% missing data overall. Likelihood analysis yielded a generally well-supported tree among families, largely consistent with trees from phylogenomic data alone. All taxa were placed in the expected families, even though 42.5% of the taxa each had >99.5% missing data, and 70.2% had >90% missing data. Our results show that missing data are not an impediment to combining ultra-large phylogenomic and supermatrix datasets and open the door to new studies that simultaneously maximize sampling of genes and taxa.
Sample:
SAMN34352650 • SRS17439209 • All experiments • All runs
Library:
Name: Pelodytes_ibericus
Instrument: Illumina HiSeq 4000
Strategy: Targeted-Capture
Source: GENOMIC
Selection: Hybrid Selection
Layout: PAIRED
Runs: 1 run, 8.5M spots, 2.4G bases, 989.3Mb
Run# of Spots# of BasesSizePublished
SRR243133698,470,0852.4G989.3Mb2023-04-26

ID:
27517376

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