Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

Standard for Training in 

Forensic DNA Sequencing 

using Capillary 

Electrophoresis

Biological Methods Subcommittee

Biology/DNA Scientific Area Committee

Organization of Scientific Area Committees (OSAC) for Forensic Science
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

OSAC Proposed Standard

Standard for Training in Forensic 

DNA Sequencing using Capillary 

Electrophoresis

Prepared by 

Biological Methods Subcommittee

Version: 1.0

Disclaimer:

This document has been developed by the Biological Methods Subcommittee of the Organization 

of Scientific Area Committees (OSAC) for Forensic Science through a consensus process and is 

proposed for further development through a Standard Developing Organization (SDO). This 

document is being made available so that the forensic science community and interested parties 

can consider the recommendations of the OSAC pertaining to applicable forensic science 

practices. The document was developed with input from experts in a broad array of forensic 

science disciplines as well as scientific research, measurement science, statistics, law, and policy. 

This document has not been published by an SDO. Its contents are subject to change during the 

standards development process. All interested groups or individuals are strongly encouraged to 

submit comments on this proposed document during the open comment period administered by 

the Academy Standards Board (www.asbstandardsboard.org). 
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

Foreword

This standard defines the minimum requirements that shall be met in a forensic DNA analyst 

training program for DNA sequencing using capillary electrophoresis (CE) methods. The aim is to 

provide a framework for quality training that will result in consistency in the forensic DNA 

community.

This standard was revised, prepared and finalized as a standard by the DNA Consensus Body of the 

AAFS Standards Board (ASB). The initial draft document was developed by the Biological Methods

Subcommittee of the Organization of Scientific Area Committees. All hyperlinks and web addresses 

shown in this document are current as of the publication date of this standard.

Keywords: Training, DNA sequencing, capillary electrophoresis
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

Table of Contents

1 Scope.......................................................................................................................................................1

2 Normative References.........................................................................................................................1

3 Terms and Definitions.........................................................................................................................1

4 Requirements .......................................................................................................................................2

4.1 Knowledge-based training..........................................................................................................2

4.2 Practical training ..........................................................................................................................3

4.3 Competency testing ......................................................................................................................3

5 Conformance...........................................................................................................................................4

Annex A .......................................................................................................................................................5
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

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1 Scope

This standard provides the requirements of a forensic DNA laboratory’s training program to ensure 

proper training in the approved methods of DNA sequencing using capillary electrophoresis (CE)

within the trainee’s forensic DNA laboratory.

2 Normative References

ASB Standard 022 - Standard for Forensic DNA Analysis Training Programs 1

3 Terms and Definitions

For the purpose of this document the following definitions apply:

3.1 

Massively parallel sequencing 

One of a number of high throughput DNA sequencing techniques. Also referred to as Next 

Generation Sequencing (NGS).

3.2 

Maxam-Gilbert sequencing

A method of DNA sequencing based on nucleobase-specific partial chemical modification of DNA 

and subsequent cleavage of the DNA backbone at sites adjacent to the modified nucleotides

3.3

Pyrosequencing

A method of DNA sequencing which is performed by detecting the nucleotide incorporated by a 

DNA polymerase

3.4

Sanger sequencing

A method of DNA sequencing for determining the order of bases in a DNA molecule based on the 

selective incorporation of chain-terminating di-deoxynucleotides by DNA polymerase during in 

vitro DNA replication. 

3.5

Sequencing

DNA sequencing is a laboratory technique used to determine the sequence of bases (A, C, G, and T) 

in a DNA molecule.

1 American Academy of Forensic Sciences Standards Board, 4200 Wisconsin Avenue, NW, Suite 106-310, 

Washington, DC 20016-2143 asb@aafs.org
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

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4 Requirements

4.1 Knowledge-based training component 

The laboratory’s training program shall provide the trainee with an understanding of the 

fundamental principles of the theory behind DNA sequencing using CE, the function of the 

sequencing reagents and CE components, the limitations of sequencing and CE, and the laboratory’s 

own DNA sequencing and CE protocols.

4.1.1. At a minimum, the knowledge-based portion of the training program shall require review of 

the following:

a) The laboratory’s protocols for DNA sequencing using CE

b) The laboratory’s applicable validation studies

c) Literature used to support validation

d) Applicable literature as assigned by the trainer (e.g., see references in Annex A)

4.1.2 At a minimum, the knowledge-based portion of the training program shall cover the 

following topics:

NOTE: Knowledge of historical methods is intended to provide an educated perspective. 

In-depth understanding of these methods may not be required for successful training.

a) Principles and limitations of sequencing methods and platforms

i. Chain termination (Sanger sequencing)

ii. Chemical (Maxam-Gilbert sequencing)

iii. High throughput (e.g.; pyrosequencing, massively parallel sequencing)

b) Quality controls used for DNA sequencing

c) Evaluation of sequencing results for controls and samples

d) Laboratory-specific CE instrumentation

i. Software

ii. Instrument maintenance and calibration 

iii. Instrument reagents/components

a. Capillary length

b. Polymer type

c. Electrophoresis conditions
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

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e) Troubleshooting

i. Sequencing failure

ii. Instrument failure

4.2 Practical training 

The laboratory’s training program shall provide the trainee with sufficient practical instruction for 

the trainee to obtain the skills for performing the DNA sequencing using CE protocols used by the 

laboratory. 

4.2.1 At a minimum, the practical portion of the training program shall include the practical 

observation of the processes at least once and until clearly understood. These include:

a) DNA sequencing and CE methods to be utilized by the trainee

b) Documentation of the process

4.2.2 At a minimum, the practical portion of the training program shall include exercises 

representative of the range, type, and complexity of routine casework and/or database 

samples processed by the laboratory. These include: 

a) DNA sequencing and CE methods to be utilized by the trainee

b) Documentation of the process

c) NOTE: The number and quality of samples processed by the trainee shall be appropriate to 

demonstrate the ability to follow the laboratory’s DNA sequencing and CE protocol(s) and 

to produce reliable and accurate results.

4.3 Competency testing 

The laboratory’s training program shall include knowledge-based and practical competency in the 

application of DNA sequencing using CE. The format of the test(s) shall meet Section 4.3 of the ASB 

Standard 022

4.3.1 Knowledge-based competency

The trainee shall successfully complete a knowledge-based test covering the critical information 

obtained during the training of DNA sequencing using CE. The test(s) shall cover, at a minimum:

a) The theoretical and scientific bases of DNA sequencing using CE

b) The function of the reagents and other components used for DNA sequencing using CE

c) The quality control steps pertaining to DNA sequencing using CE

d) The laboratory’s analytical procedures pertaining to DNA sequencing using CE methods

4.3.2 Practical competency
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

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The trainee shall successfully complete a practical test covering each of the DNA sequencing using

CE protocol(s) for which he or she will be independently authorized. The trainee shall be able to 

satisfactorily perform the following, as applicable:

a) Properly and accurately execute the analytical procedures related to DNA sequencing using 

CE

b) Apply the laboratory’s analytical procedures to a set of samples representing the range of 

DNA quality and quantity expected to be encountered in the laboratory

c) Operate relevant equipment and instrumentation used in the laboratory

d) Document work performed in accordance with laboratory procedures

5 Conformance

In order to demonstrate conformance with this standard, the laboratory shall meet Section 5 of the 

ASB Standard 022.
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

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Annex A

(informative)

Bibliography

The following information provides a list of the literature resources that may assist the DNA 

technical leader in defining the breadth and scope of the materials to be reviewed by the trainee. 

This list is not meant to be all inclusive. The laboratory shall develop a list tailored to its specific 

needs. Updated references shall be added to the laboratory’s list as new methods or technologies 

are incorporated into the laboratory’s protocols. 

1) Berger, C., & Parson, W. (2009). Mini-midi-mito: Adapting the amplification and sequencing 

strategy of mtDNA to the degradation state of crime science samples. Forensic Science 

International: Genetics, 3, 96-103.

2) Butler, J. M. (2012). Advanced Topics in Forensic DNA Typing: Methodology. Academic Press, 

chapter 14, 405-456.

3) Davis C et al. Sequencing the hypervariable regions of human mitochondrial DNA using 

massively parallel sequencing: Enhanced data acquisition for DNA samples encountered in 

forensic testing, Legal Med. 2015; 17:123–7.

4) Holland, MM & Parsons TJ. (1999). Mitochondrial DNA sequence analysis – Validation and use 

for forensic casework. Forensic Science Review. 11: 21-50.

5) Lee, L.G. et al (1997). New energy transfer dyes for DNA sequencing. Nucleic Acids Research, 25, 

2816-2822.

6) Maxam, A. M., and Gilbert, W. (1977) A new method for sequencing DNA Proc. Natl. Acad. Sci. 

USA 74, 560–564.

7) Parson W. et al. (2013). DNA Commission of the International Society for Forensic Genetics: 

revised and extended guidelines for mitochondrial DNA typing. Forensic Science International –

Genetics. 13: 134-142.

8) Rasmussen, E. M., et al. (2002). Sequencing strategy of mitochondrial HV1 and HV2 DNA with 

length heteroplasmy. Forensic Science International, 129, 209-213.

9) Sanger, F., et. al. (1977). DNA sequencing with chain-terminating inhibitors. Proceedings of the 

National Academy of Sciences of the United States of America, 74, 5463-5467.

10) Stewart, J.E. et al. (2003). Evaluation of a multipcapillary electrophoresis instrument for 

mitochondrial DNA typing. Journal of Forensic Sciences, 48, 571-580.
Standard for Training in Forensic DNA Sequencing using Capillary 

Electrophoresis 

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11)Tully, G., et al. (2001). Considerations of the European DNA Profiling (EDNAP) group on the 

working practices, nomenclature and interpretation of mitochondrial DNA profiles. Forensic 

Science International. 124: 83-91.

12)Wilson, M.R. et al. (1995). Validation of mitochondrial sequencing for forensic casework 

analysis. Int J Legal Medicine. 108:68-74.