2021-N-0001 Wildlife Forensics MethodCollection of Known DNA Samples from Domestic Mammals

OSAC 2021-N-0001

Wildlife Forensics MethodCollection of Known DNA

Samples from Domestic

Mammals

Wildlife Forensic Biology

Biology Scientific Area Committee

Organization of Scientific Area Committees (OSAC) for Forensic Science
2021-N-0001 Wildlife Forensics MethodCollection of Known DNA Samples from Domestic Mammals

OSAC Proposed Standard

OSAC 2021-N-0001

Wildlife Forensics Method-Collection of

Known DNA Samples from Domestic

Mammals

Prepared by

Wildlife Forensics Biology Subcommittee

Version: 1.1

March 3, 2021

Disclaimer:

This OSAC Proposed Standard was written by the Wildlife Forensic Biology of the Organization of

Scientific Area Committees (OSAC) for Forensic Science following a process that includes an open

comment period. This Proposed Standard will be submitted to a standards developing organization

and is subject to change.

There may be references in an OSAC Proposed Standard to other publications under development

by OSAC. The information in the Proposed Standard, and underlying concepts and methodologies,

may be used by the forensic-science community before the completion of such companion

publications.

Any identification of commercial equipment, instruments, or materials in the Proposed Standard is

not a recommendation or endorsement by the U.S. Government and does not imply that the

equipment, instruments, or materials are necessarily the best available for the purpose.
2021-N-0001 Wildlife Forensics MethodCollection of Known DNA Samples from Domestic Mammals

Wildlife Forensics Method-Collection of Known DNA

Samples from Domestic Mammals

Foreword

This standard presents protocols for collecting DNA samples for use in genetic analysis of

domestic mammals. It is based on a draft from the Wildlife Forensic Biology of the

Organization of Scientific Area Committees (OSAC) for Forensic Science.

Keywords: DNA sample collection, domestic mammal, dog, cat, horse, cattle, pig, sheep
2021-N-0001 Wildlife Forensics MethodCollection of Known DNA Samples from Domestic Mammals

1 Scope

This standard provides the protocol for obtaining genetic known evidence samples (buccal swabs and

pulled hair) for the purpose of genetic analysis of domestic mammals. This standard does not address

sampling of wild animals.

2 Normative Reference

There are no normative reference documents. Annex A, Bibliography, contains informative references.

3 Terms and Definitions

3.1

Buccal swab

A cotton swab or similar collection substrate; used in a relatively non-invasive sample collection

technique for scraping the inside of a mouth to collect cells from the inner cheek lining; this is a

common method for collecting and preserving samples for DNA testing from known individuals.

3.2

chain of custody

Chronological record of the handling and storage of an item from its point of collection to its

final return or disposal.

3.3

known sample

Biological material for which the identity of the donor is established and used for comparison

purposes.

3.5

nuclear DNA

The DNA inside a cell’s nucleus, existing in the form of chromosomes.

4 Requirements

4.1 General

4.1.1 All known evidence samples submitted shall have a unique name or number.

4.1.2 All required paperwork (e.g. submission forms, chain of custody forms) shall be completed.

4.1.3 Pertinent information, such as sample collector, collection date and site, sample type (e.g., buccal

swab, hairs), swab type, sample storage conditions, etc., should be recorded.

4.2 Buccal swabs

4.2.1 Buccal swabbing using cotton, nylon, or other swab types is the preferred method for collecting

DNA from domestic cats and dogs. Buccal swabs should be sterile.
2021-N-0001 Wildlife Forensics MethodCollection of Known DNA Samples from Domestic Mammals

NOTE: While many swab types are acceptable for buccal swabbing, cotton swabs can sometimes be

problematic for buccal collection from dogs.

4.2.2 Preparing to collect buccal swabs

a) Isolate any animal(s) to be tested from other individuals (including nursing

animals) and remove food and water at least 30 minutes prior to sample collection.

b) If buccal swabs from more than one animal will be collected, complete the process

for one animal and change gloves or clean hands before collecting from the next.

c) Use at least 2 buccal swabs per animal to obtain replicate samples.

d) Prepare a paper envelope or other container for each animal. Label it so as to

uniquely identify the animal from which the sample is collected.

NOTE: Other packaging options may be appropriate as long as buccal swabs are stored in a way that

minimizes mold, bacterial growth, and sample degradation.

4.2.3 Collecting the sample

a) Open the swab packaging and remove the swab by its handle.

b) Place the head of the swab against the inside of the cheek and gums, and swirl/wipe

vigorously 8–10 times.

c) Allow the swab to dry in a clean environment, unless cells/swabs are directly transferred to a

different storage medium (e.g. preservative liquid, preservative paper).

d) Place the dry swabs in the labeled envelope (or equivalent). Seal the envelope and sign and

date the seal.

NOTE: Dry swabs may be stable at room temperature, however refrigeration is recommended for short

term storage (days to weeks) while freezing is recommended for longer term storage.

4.3 Pulled hair

4.3.1 Pulled hairs with roots are the preferred DNA source for domestic mammals other than cats and

dogs. Because nuclear DNA is only present in high quantities in the root of the hair, cut hairs are

not acceptable.

4.3.2 Preparing to collect hairs

a) If hairs from more than one animal will be collected, complete the process for one animal and

change gloves or clean hands (and pliers or similar if used) before collecting hairs from the

next.

b) Prepare a paper envelope or other container for each animal. Label it so as to uniquely identify

the animal from which the sample is collected.

NOTE: Other packaging options may be appropriate as long as hairs are stored in a way that minimizes

mold, bacterial growth, and sample degradation.
2021-N-0001 Wildlife Forensics MethodCollection of Known DNA Samples from Domestic Mammals

4.3.3 Collecting the sample

a) Collect thick hairs such as those from the mane, tail, or fetlock (base of leg just above the

hoof). The area on the animal from which hairs are collected should be dry. If dirt or debris is

present, brush the area to remove it.

b) Use fingers or pliers (or similar) to grasp hair close to the skin and pull approximately 10

hairs at once (do not cut hairs). Repeat this until 20–30 hairs with roots have been obtained.

NOTE: For pigs, which have very thick hairs, 5–10 hairs with roots are sufficient.

c) Place the hairs in the labeled envelope (or equivalent). Seal the envelope and sign and date the

seal.

NOTE: Hairs may be stable at room temperature if kept dry and free from pests (e.g. dermestid beetles), however

refrigerated or frozen are also acceptable storage conditions.
2021-N-0001 Wildlife Forensics MethodCollection of Known DNA Samples from Domestic Mammals

Annex A

(informative)

Bibliography

1. Ellegren H, Johansson M, Sandberg K, Andersson L, Cloning of highly polymorphic

microsatellites in the horse, Animal Genetics. 1991, vol. 23, 133–142.

2. Irion DN, Schaffer AL, Famula TR, Eggleston ML, Hughes SS, Pedersen NC, Analysis of Genetic

Variation in 28 Dog Breed Populations With 100 Microsatellite Markers, Journal of Heredity.

2003, vol. 94, 81–87. https://doi.org/10.1093/jhered/esg004

3. Metallinos DL. Canine Molecular genetic testing. Veterinary Clinics: Small Animal Practice. 2001,

vol. 31, 421–431.

4. Oberbauer AM, Grossman DI, Eggleston ML, Irion DN, Schaffer AL, Pedersen NC, Belanger JM.

Alternatives to blood as a source of DNA for large-scale scanning studies of canine genome

linkages. Veterinary Research Communications. 2003, vol. 27, 27–38.