scope:

The intended users of this guideline are clinical laboratories involved in the development, validation, verification, and implementation of sequencing-based assays.

This guideline specifies recommendations for the sequencing process, including specimen collection and handling, isolation of nucleic acid, amplification and sequencing of nucleic acids, and general interpretation and reporting of genotyping results. It is the intent of this document to provide instruction for verifying that the sequence obtained is accurate and suitable for subsequent interpretation; to address general interpretation of the sequence; and to provide QA/QC considerations for each step of the process, as appropriate. It is also intended to assist laboratories in generating appropriate and efficient validation across sequencing methods and applications. Sanger-based DNA sequencing and general aspects of massively parallel sequencing (MPS) are addressed in this guideline with specific examples.

This guideline:

• Does not comprehensively address platform-specific issues, because sequencing technology is rapidly evolving

• Provides general guidance for interpreting sequencing results and does not address the medical interpretation for a given patient, which is under the purview of the health care provider

• Is relevant to germline, somatic, and microbiological applications in clinical settings