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. 2020 Nov;32(6):767-775.
doi: 10.1177/1040638720905833. Epub 2020 Feb 18.

Guidelines for Sanger sequencing and molecular assay monitoring

Beate M Crossley  1   2   3   4   5   6 , Jianfa Bai  1   2   3   4   5   6 , Amy Glaser  1   2   3   4   5   6 , Roger Maes  1   2   3   4   5   6 , Elizabeth Porter  1   2   3   4   5   6 , Mary Lea Killian  1   2   3   4   5   6 , Travis Clement  1   2   3   4   5   6 , Kathy Toohey-Kurth  1   2   3   4   5   6
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Guidelines for Sanger sequencing and molecular assay monitoring

Beate M Crossley et al. J Vet Diagn Invest. 2020 Nov.

Abstract

Genetic sequencing, or DNA sequencing, using the Sanger technique has become widely used in the veterinary diagnostic community. This technology plays a role in verification of PCR results and is used to provide the genetic sequence data needed for phylogenetic analysis, epidemiologic studies, and forensic investigations. The Laboratory Technology Committee of the American Association of Veterinary Laboratory Diagnosticians has prepared guidelines for sample preparation, submission to sequencing facilities or instrumentation, quality assessment of nucleic acid sequence data performed, and for generating basic sequencing data and phylogenetic analysis for diagnostic applications. This guidance is aimed at assisting laboratories in providing consistent, high-quality, and reliable sequence data when using Sanger-based genetic sequencing as a component of their laboratory services.

Keywords: Sanger sequencing; quality score; validation.

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Conflict of interest statement

Declaration of conflicting interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
Examples of sequences. A. A good nucleic acid sequence (porcine circovirus 2). B. Dual infections are indicated by overlapping peaks, example pointed out by arrow (infectious bronchitis virus). C. High background noise (avian influenza virus), using FinchTV software v.1.4.0.
See this image and copyright information in PMC

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