Guidelines for Sanger sequencing and molecular assay monitoring
- PMID: 32070230
- PMCID: PMC7649556
- DOI: 10.1177/1040638720905833
Guidelines for Sanger sequencing and molecular assay monitoring
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.
Conflict of interest statement
Figures
References
-
- Altschul SF, et al. Basic local alignment search tool. J Mol Biol 1990;215:403–410. - PubMed
-
- Campos P, Gilbert TP. DNA extraction from formalin-fixed material. Methods Mol Biol 2012;840:81–85. - PubMed
-
- Do H, Dobrovic A. Sequence artifacts in DNA from formalin-fixed tissues: causes and strategies for minimization. Clin Chem 2015;61:64–71. - PubMed
-
- Downey N. Extraction of DNA from agarose gels. In: Casali N, Preston A, eds. E. coli Plasmid Vectors: Methods and Applications. (Vol. 235, Methods in Molecular Biology series.) Totowa, NJ: Humana Press, 2003:137–139. - PubMed