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1.
Med Sci Law ; 57(1): 7-11, 2017 Jan.
Article in English | MEDLINE | ID: mdl-27794077

ABSTRACT

The purpose of this study was to evaluate which DNA extraction method yields the highest quantity of DNA from chewing gum. In this study, several popular extraction methods were tested, including Chelex-100, phenol-chloroform-isoamyl alcohol (PCIA), DNA IQ, PrepFiler, and QIAamp Investigator, and the quantity of DNA recovered from chewing gum was determined using real-time polymerase chain reaction with Quantifiler. Chewed gum control samples were submitted by anonymous healthy adult donors, and discarded environmental chewing gum samples simulating forensic evidence were collected from outside public areas (e.g., campus bus stops, streets, and sidewalks). As expected, results indicate that all methods tested yielded sufficient amplifiable human DNA from chewing gum using the wet-swab method. The QIAamp performed best when DNA was extracted from whole pieces of control gum (142.7 ng on average), and the DNA IQ method performed best on the environmental whole gum samples (29.0 ng on average). On average, the QIAamp kit also recovered the most DNA from saliva swabs. The PCIA method demonstrated the highest yield with wet swabs of the environmental gum (26.4 ng of DNA on average). However, this method should be avoided with whole gum samples (no DNA yield) due to the action of the organic reagents in dissolving and softening the gum and inhibiting DNA recovery during the extraction.


Subject(s)
Chewing Gum , DNA Fingerprinting/methods , DNA/isolation & purification , Humans , Real-Time Polymerase Chain Reaction
2.
Anal Biochem ; 478: 128-30, 2015 Jun 01.
Article in English | MEDLINE | ID: mdl-25197027

ABSTRACT

We compared four proteases in the QIAamp DNA Investigator Kit (Qiagen) to extract DNA for use in multiplex polymerase chain reaction (PCR) assays. The aim was to evaluate alternate proteases for improved DNA recovery as compared with proteinase K for forensic, biochemical research, genetic paternity and immigration, and molecular diagnostic purposes. The Quantifiler Kit TaqMan quantitative PCR assay was used to measure the recovery of DNA from human blood, semen, buccal cells, breastmilk, and earwax in addition to low-template samples, including diluted samples, computer keyboard swabs, chewing gum, and cigarette butts. All methods yielded amplifiable DNA from all samples.


Subject(s)
DNA/isolation & purification , DNA/metabolism , Peptide Hydrolases/metabolism , Polymerase Chain Reaction/methods , Ananas/enzymology , Bromelains/metabolism , Carica/enzymology , DNA/analysis , DNA/blood , Endopeptidase K/metabolism , Fungi/enzymology , Humans , Papain/metabolism , Reagent Kits, Diagnostic
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