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Department of Forensic Biology

Forensic Biology Protocols for Mitochondrial DNA Analysis Manuals

This manual contains individual procedures pertaining to Mitochondrial DNA testing of biological specimens.

Procedures on this page are effective as of June 20, 2016 and later. The dates effective are shown as MM/DD/YY - MM/DD/YY in the file name.

Note: The most current manual in each section is the first link listed.

Click or press the enter key on any topic below to access the relevant PDF file of the associated procedure.

Pre-Amplification Procedures

Mitochondrial DNA Guidelines

Hair Evidence Examination

Washing Hair for Mitochondrial or Nuclear DNA Testing

Organic Extraction for Mitochondrial or Nuclear DNA Testing

mitoMPS Massively Parallel Sequencing Procedures

Mitochondrial DNA Guidelines for mitoMPS casework

Library Amplification with the PowerSeq CRM kit

Ampure XP bead-based library purification

Library Quantification with the PowerSeq Quant MS kit

Normalization and Pooling of MPS Libraries

mtDNA Sequencing using the MiSeq/MiSeq FGx

mitoMPS Analysis using the Qiagen CLC Workbench and AQME Toolkit

Interpretation Guidelines for mtDNA MPS

Statistical Analysis for Massively Parallel Sequencing

Sanger Sequencing Procedures

Duplex Mitochondrial DNA PCR Amplification

Quantitation using Agilent 2100 Bioanalyzer

Cycle-Sequencing

As of 8/23/21 the Cycle – Sequencing procedure was incorporated into the combined procedure named “Mito Cycle Sequencing, Cleanup, and 3130xl Sequencing".

SDS Cleanup

As of 8/23/21 the SDS Cleanup procedure was incorporated into the combined procedure named “Mito Cycle Sequencing, Cleanup, and 3130xl Sequencing”.

ExoSAP-IT Sample Cleanup

Centri-Sep Sample Filtration

As of 8/23/21 the Centri-Sep Sample Filtration procedure was incorporated into the combined procedure named “Mito Cycle Sequencing, Cleanup, and 3130xl Sequencing”.

ABI 3130xl Sequencing

As of 8/23/21 the ABI 3130xl Sequencing procedure was incorporated into the combined procedure named “Mito Cycle Sequencing, Cleanup, and 3130xl Sequencing”.

Mito Cycle Sequencing, Cleanup, and 3130xl Sequencing

Mitochondrial DNA Sequencing Analysis

Editing Guidelines

Sequence Nomenclature and Alignment

Interpretation Guidelines

Statistical Analysis

Creation of a Casefile CD

References

Preparation of Phase Lock Gel (PLG) Tubes

This procedure has been retired as of 07/09/19.

Appendix A - Oligonucleotide Primer Sequences

Appendix B - Mitochondrial DNA Primer Locations

Appendix C - Revised Cambridge Reference Sequence