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SRX2491926: genome sequencing of Sindbis virus experimental population
1 ILLUMINA (Illumina HiSeq 2500) run: 449,411 spots, 68.3M bases, 39.9Mb downloads

Design: Viral RNA was isolated using the QIAamp Viral RNA mini kit (Qiagen). All subsequent steps, from reverse transcription through sequencing, were completed in duplicate, resulting in two technical replicates of each sample (entered here as ÔAŐ and ÔBŐ). cDNA was generated by reverse-transcription with Superscript II (Life Technologies) using random hexamer primers. The entire genome sequence was amplified via PCR with GoTaq DNA Polymerase (Promega) using eight primer pairs, generating overlapping PCR fragments 1.5-2.1 kb in length. Amplified genome fragments were purified using the QIAquick PCR Purification Kit (Qiagen), and the eight fragments from each virus sample were combined to create an equamolar mixture. Libraries were prepared from the pooled viral amplicons using the Nextera XT Kit (Illumina Inc.) and the Nextera Index Kit (Illumina Inc.). Samples were sequenced in a single lane via paired-end, 75-bp read Illumina HiSeq 2500 (Illumina Inc.) at the Yale Center for Genome Analysis.
Submitted by: Yale University
Study: Sindbis virus Raw sequence reads
show Abstracthide Abstract
Understanding the dynamics of molecular evolution is a fundamental goal of evolutionary biology. Experimental evolution in microbial populations provides a tractable model in which to study these dynamics. In this study, we used next-generation sequencing to map adaptive walks in experimentally evolved Sindbis virus (SINV) populations. Virus populations were allowed to evolve in one of two tissue culture treatments: an environment in which they were shifted suddenly onto a novel host cell type and propagated on solely the novel host, or in a mixed-cell environment where the prevalence of the novel host increased gradually over time. Virus populations were sampled at time points over the course of their evolution, and whole-genome population-level sequencing was used to track allele frequencies at each time point. This produces a high-resolution map of how new alleles enter the populations and change in frequency over time. These data were used to answer questions related to how the rate of environmental change affects the distribution of fixed mutational effect sizes, the degree of clonal interference, and the number of mutations that fix in each fixation event.
Sample:
SAMN06220896 • SRS1921815 • All experiments • All runs
Organism: Sindbis virus
Library:
Name: Population19_Passage22_B
Instrument: Illumina HiSeq 2500
Strategy: AMPLICON
Source: VIRAL RNA
Selection: RT-PCR
Layout: PAIRED
Runs: 1 run, 449,411 spots, 68.3M bases, 39.9Mb
Run# of Spots# of BasesSizePublished
SRR5175663449,41168.3M39.9Mb2017-01-18

ID:
3612978

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