AOAC SPADA VNGS - Final

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(2) Artefacts.— No artefacts found in final sequence (40, 41, 42).

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(3) Sequencing platform specific error profiles .—All platform associated errors should be resolved

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(43, 44).

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(4) Variation in quality scores across the sequence read.—Sequence reads shall have an overall

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resolved Q score >20 (45).

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(5) Biases in sequence data driven by base composition.— GC-rich sequence bias shall be anticipated

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and resolved, based on species specificity or nucleic acid repair (46, 47).

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(6) Departure from suboptimal library fragment sizes . — If possible, average library fragment size

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shall be provided as metadata (48).

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(7) Contamination from known and unknown species other than the sequencing target .—

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Contaminating species sequences shall be removed (49).

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(8) Insert size .—Insert size and type of library preparation should be provided.

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(9) Number of reads.— The minimum read depth is 20X.

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(10) Base calling.— Base calling protocol should be provided.

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(11) Sequence length distribution.— Library quality should be recorded.

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(12) Length of longest contig.— Length of longest contig can be provided.

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(13) N50 .—N50 should be given in annotations (50).

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(14) NG50 .—NG50 should be given in annotations (50).

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(15) Number of contigs.— Number of contigs should be given in annotations.

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(16) Base composition.— The proportions of the four bases (adenine, cytosine, guanine, and thymine

or uracil) present in DNA or RNA expressed as the percentage (mol %) of G plus C should be given in 233

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annotations.

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(17) Coverage .—At the run level, minimum 20X. At the sample level, it depends on the application.

AOAC Draft Standard – Version 09282022; Public Comment Revisions

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