Selected article for: "copy number and PCR product"
Author: William Fitzsimmons; Robert J. Woods; John T. McCrone; Andrew Woodman; Jamie J. Arnold; Madhumita Yennawar; Richard Evans; Craig E. Cameron; Adam S. Lauring
Title: A speed-fidelity trade-off determines the mutation rate and virulence of an RNA virus
  • Document date: 2018_4_27
    • ID: 8p24gszj_71
    Snippet: For tissue distribution analyses, mice were infected intravenously via tail vein with 100µl and 499 observed twice daily as above. Mice were euthanized for severe morbidity or on day 5, the The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/309880 doi: bioRxiv preprint 21 for 5 min, then 4°C hold. For each sample, amplification of all 4 fragments was confirmed by gel 520 elect.....
    Document: For tissue distribution analyses, mice were infected intravenously via tail vein with 100µl and 499 observed twice daily as above. Mice were euthanized for severe morbidity or on day 5, the The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/309880 doi: bioRxiv preprint 21 for 5 min, then 4°C hold. For each sample, amplification of all 4 fragments was confirmed by gel 520 electrophoresis and equal quantities of each PCR product were pooled. Seven hundred fifty 521 nanograms of each cDNA mixture were sheared to an average size of 300 to 400bp using a 522 Covaris S220 focused ultrasonicator. Sequencing libraries were prepared using the NEBNext The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/309880 doi: bioRxiv preprint 22 separate RT-PCR reactions and sequencing libraries for samples with copy number 10 3 -10 5 546 genomes per µl (for example in tissue studies). Our strategy for variant calling as well as our 547 benchmarked sensitivity and specificity are described in [46] and all code can be found at 548 https://github.com/lauringlab/variant_pipeline.

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