Percentage of errors by type of acquired diversity determined during sample preparation.

<p>(A) Percentage of error attributed to synonymous vs. non-synonymous variants. (B) Percentage of errors attributed to transitions vs. transversions. (C) Percentage of errors attributed to insertions or deletions vs. Intra-host single nucleotide variants (iSNV).</p

Sources of error per sample preparation method.

<p>(A) Intra-host single nucleotide variants (iSNVs) that were present in multiple samples at greater than 1% of population are shown in a heat map format to visualize patterned diversity acquired during sample preparation. The total number of samples containing iSNVs in greater than 1% of population are summarized in the column “#> 1% of Pop” in green. “Codon” column (second column from right) provides both the nucleotide and protein translation of the site. (B) The detected mean error rates of iSNVs greater than 0. 2% of population (error/site/copy) are stratified by presence in the plasmid (Origin), detection after transcription/reverse transcription (Transc) or preparation, and preparation/sequencer error (Prep). (C) Error profiles are expressed as the number of iSNVs per percent of population obtained from each of the 5 sample preparation methods.</p

Study sample diagram.

<p>Study sample diagram.</p

Total error by sample preparation method.

<p>(A) The mean read depth per position from two DNA controls (PLASMID and P_AMP) and five RNA sample preparation methods (n = 2 of replicate experiments). (B) Errors calculated as error/site/copy (plasmid or transcript) are presented as the average of duplicate experiments as in A. Intra-host single nucleotide variants present in the original plasmid were removed from the calculation of the error (reference positions: 4,697 and 4,725). Student t-tests were performed to demonstrate differences between the mean error per site per copy in the control plasmid (* = <i>p</i><0.05) and each sample preparation method. (C) The error calculated in B is converted to fold change over the control (“PLASMID”). Error bars in all panels represent standard deviation of the mean.</p

Intrahost single nucleotide variants (iSNVs) of EBOV/Kik-9510621 “R4415” (passage 3) comprising ≥2% of total population as compared to “134” (GenBank #AY354458).

<p>Intrahost single nucleotide variants (iSNVs) of EBOV/Kik-9510621 “R4415” (passage 3) comprising ≥2% of total population as compared to “134” (GenBank #AY354458).</p

Intrahost single nucleotide variants (iSNVs) of EBOV/Kik-9510621 “R4368” (passage 4) comprising ≥2% of total population as compared to “134” (GenBank #AY354458).

<p>Intrahost single nucleotide variants (iSNVs) of EBOV/Kik-9510621 “R4368” (passage 4) comprising ≥2% of total population as compared to “134” (GenBank #AY354458).</p

<i>GP</i> gene editing site composition in EBOV/Kik-9510621 NHP challenge stocks.

<p><i>GP</i> gene editing site composition in EBOV/Kik-9510621 NHP challenge stocks.</p

History of Ebola virus variant Kikwit (isolate 9510621) challenge stocks used at the United States Army Medical Research Institute of Infectious Diseases (USAMRIID), CDC, Centers for Disease Control and Prevention; NHP, nonhuman primate; SPBLOG, Special Pathogens Branch Log; UTMB, University of Texas Medical Branch; VSP, virus seed pool.

<p>History of Ebola virus variant Kikwit (isolate 9510621) challenge stocks used at the United States Army Medical Research Institute of Infectious Diseases (USAMRIID), CDC, Centers for Disease Control and Prevention; NHP, nonhuman primate; SPBLOG, Special Pathogens Branch Log; UTMB, University of Texas Medical Branch; VSP, virus seed pool.</p