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Sara Rowley

Terry Roylance

CHEM 1010

17 November 2022

DNA Testing and Typing in Forensics

Biological specimens have been used as evidence in forensic cases since the 1980’s.

Since this time multiple developments have taken place allowing forensic scientists to test

smaller samples with greater accuracy than ever before. DNA fingerprinting is an outgrowth of

its use in a medical context, originally developed to analyze disease-causing genes based on

comparison between the patient's DNA and DNA from their relatives to study inheritance

patterns. It’s important to note that forensic specimens are inherently less reliable than medical

ones. They are often too small to accommodate repeat testing, come from multiple sources, and

are contaminated and/or degraded. Consequently, specific instrumentation has been developed to

carry out the majority of sample processing and improved chemistries have been employed to

allow for easier interpretation. The four main steps of DNA testing include extraction,

quantitation, amplification and capillary electrophoresis. DNA testing creates a fingerprint that

excludes all other individuals except one within a certain percentage. It can be used to identify

victims, suspects, and establish leads in criminal cases.

Extraction is a term which describes the process used to pull DNA molecules from the

nucleus of the cell into solution, and separate them from other cellular material that may be

present (known as inhibitors). Extraction can be performed manually or with the use of

specialized robotic instruments. It’s important to have a thorough understanding of the sample

and its inhibitors when deciding which method of extraction should be utilized.
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After the DNA is extracted, the quality and quantity of the DNA must be evaluated to

determine that the sample comes from a human source, and that there is enough sample to have

the DNA amplified. The two common options for quantification both utilize optical technology

to quantify nucleic acids. UV-Vis measurement uses the photometric measurement of the nucleic

acid based on its intrinsic absorptive properties. Fluorescence measurement uses fluorogenic

dyes that only bond with DNA and RNA. Amplification is completed via a process known as

Polymerase Chain Reaction (PCR), where short synthetic DNA fragments called primers are

used to select specific sequences of DNA to be amplified. Multiple rounds of DNA synthesis

take place and millions of copies of that DNA sequence are produced.

After the sample’s PCR reaction is complete, there is a mixture of amplified DNA

molecules which must then be divided using capillary electrophoresis (CE). The instrument used

to complete CE has a thin capillary filled with a gel-like polymer and a positively charged anode

at the end of the chain. An electrical current is applied to the negatively charged DNA sample

which travels along the capillary chain to seek a positively charged anode. The speed with which

each molecule travels is dependent on its size, thus separating them accordingly. This data is

collected by a computer attached to the CE instrument, and turned into a DNA profile by a

software program.

After CE the DNA fingerprint will appear in the form of a band, often likened to a

barcode. The technician analyzing the DNA pattern must first determine that there are an

appropriate number of bands present. This is especially important because band number

discrepancies can arise when multiple DNA fingerprints are present, or if the sample is degraded.

The pattern should always be compared to a control pattern of known DNA to ensure that the

hybridization was performed correctly.

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DNA typing is commonly profiled by short tandem repeat (STR) loci. Based on STR loci

there are two common practices for identification. The best method is via direct matching, where

the reference DNA is obtained from a victim or their belongings. Kinship testing is another

commonly used method when direct matching is impossible. Using this method, DNA from a

relative is collected and a probability distribution of genotypes is used to infer the victim’s

identity. Except in cases where DNA evidence is used to exclude a suspect, the STR must be

compared against a wide range of possibilities, and the results require statistical analysis of

population frequency to determine how accurate the match is with a likelihood ratio (LR). For a

specimen to be considered a match to a reference sample, the LR must be precise, objective, and

applied uniformly, otherwise the result must be considered “inconclusive'' or a “nonmatch”.

In conclusion, DNA typing was developed in the 1980’s to identify individuals using

specimens that are too small, come from multiple sources, and are contaminated and/or

degraded. Forensic scientists use four main steps to test DNA. They extract DNA molecules

from other cellular materials. They quantify the specimens, then they are amplified. Finally the

DNA molecules undergo capillary electrophoresis to create a DNA fingerprint that appears in the

form of a band, which will then be analyzed and compared to reference DNA to match victims

and exclude suspects.

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Bibliography

Bureau of Criminal Apprehension. “DNA TESTING PROCEDURES.” Bureau of Criminal

Apprehension: A Division of the Minnesota Department of Public Safety, 2022,

dps.mn.gov/divisions/bca/bca-divisions/forensic-science/Pages/dna-

procedures.aspx#:~:text=The%20DNA%20testing%20process%20is,%2C

%20amplification%2C%20and%20capillary%20electrophoresis.

Moreno, Lilliana I. and Florida International University. The Effect of Sample and Sample

Matrix on DNA Processing: Mechanisms for the Detection and Management of

Inhibition in Forensic Samples. 23 Mar. 2015,

digitalcommons.fiu.edu/cgi/viewcontent.cgi?article=3020&context=etd.

Narahara, Maiko, et al. “Application of Permanents of Square Matrices for DNA Identification in

Multiple-fatality Cases - BMC Genomic Data.” BioMed Central, 21 Aug. 2013,

bmcgenomdata.biomedcentral.com/articles/10.1186/1471-2156-14-72.

National Research Council (US) Committee on DNA Technology in Forensic Science. “DNA

Technology in Forensic Science.” National Library of Medicine; National Center for

Biotechnology Information, 1992, www.ncbi.nlm.nih.gov/books/NBK234539/#:~:text=

Forensic%20DNA%20typing%20often%20involves,lacks%20built%2Din

%20consistency%20checks

Steendam, Kathleen Van, et al. “Mass Spectrometry-based Proteomics as a Tool to Identify

Biological Matrices in Forensic Science.” National Library of Medicine; National Center

for Biotechnology Information, Mar. 2013, pubmed.ncbi.nlm.nih.gov/22843116.

ThermoFisher Scientific. “RNA/DNA Quantification.” ThermoFisher Scientific, 2022,

www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/nucleic-
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