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That so many sequences contained sequencing errors in the primers (n = 377) was surprising, and warranted further investigation.
Over 5,000,000 Illumina reads were generated from two replicate single-cell amplified genomes (SAGs), but neither SAG contained sequencing reads matching the contaminating DNA.
Several of the ORFs in Table 1 contained sequencing errors or were identified as fragments and were manually corrected/assembled (see Methods for details).
For three of the taxa available on GenBank, some genes likely contained sequencing errors, as stop codons were present within the genes.
This was corroborated by comparing raw sequences; While both the reference mapping and the de novo assembly contained sequencing errors, these errors influenced the consensus of reference-mapped reads more severely.
After alkylation, the reduction of proteins was repeated, and they were digested in a buffer (50 mM ammonium bicarbonate, 1 mM calcium chloride) that contained sequencing grade modified trypsin (Promega), at a sample protein to trypsin ratio of 25:1, at 37 °C with gentle shaking overnight.
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The three-gene dataset contained sequences from 70 specimens.
The 1976 movie, All the President's Men, starring Robert Redford as Woodward and Dustin Hoffman as Bernstein, contained sequences showing Woodward meeting Felt.
The families that contained sequences that have multiple functional annotations were also removed (Supplementary Tables 4.1 and 4.2).
We've pushed our mediums to the limit - especially with After Dido, which contained sequences I wasn't 100% thrilled with.
Contig_B (279,442 bp) contained sequences of the S4 and F1 ORFs (Additional file 1: Figure S1).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com