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DNA transfer within forensic exhibit packaging: Potential for DNA loss and relocation

FSI Genetics, 2012

Study Design

Addressed Question

DNA transfer within forensic exhibit packaging under controlled conditions

Activity Context

Professional

Category

PersistenceTransfer Scenario

Specifications

Persistence with Packaging and TransportSurfaceTransfer SequenceTransfer via Vector

Variables of Interest

fit of the packagingtype of exhibitweightvolume of saliva

Stringency of Control

Controlled

Number of Individuals

1

Replicates per Individual and Condition

4

Nucleic Acid

DNA

Bodily Origin

saliva

Depositor & Contact

Depositor Characteristics

N/A

Criteria for Shedder Status

N/A

Previous Activities

N/A

Contact Scenario

transfer of saliva on designated are of exhibit - drying time (24 h) - packaging - carrying - shaking on orbital mixer (1 h, 150 rpm) - transport -sampling

Primary Substrate

Primary Substrate Type

4x4 cm deposit area of cotton fabric or plastic

Primary Substrate Material

CottonPlastic

Deposit

5 or 50 µl saliva

Delay

N/A

Secondary Substrate

Secondary Substrate Type

mock exhibit or plastic exhibit bag

Secondary Substrate Material

CottonPlastic

Secondary Substrate Contact

pressure and friction 1h

Further Transfer

N/A

Sampling

Background DNA on Sampled Surface

Negative (Confirmed)

Sampling Time

delayed (by contact scenario)

Persistence

packaging

Sampling Method

direct extraction from all surfaces

Sampling Area

7.5cmx4cm deposit squares, whole bag surface

Laboratory Analysis

Extraction

5% Chelex, Amicon Ultra YM-30 centrifugal filter (Millipore)

DNA Quantification

Quantifiler Human DNA Quantification kit (Applied biosystems), ABI PRISM 7500 SDS Instrument

Input for Profiling

N/A

Profiling

only for the replicate with the highest conc.: AmpFlSTR Profiler plus (Applied Biosystems)

Reference Samples

taken from depositor

Profile Interpretation and Mixture Analysis

confirmation of single source profile from saliva depositor

RNA Data Interpretation

N/A

Results

DNA Quantity

n.s. (only transfer rates in % calculated from conc. shown)

Profile Quality

single full or partial profile from depositor

Parameter Used for Comparison

% transfer = DNA on specific surface (ng)/(DNA on all surfaces (ng))

Summary of Results

DNA transfer from deposit area ranges from 0-100% (avg. 43.6%) to all areas of the exhibit packaging and the substrate; many variables influence DNA transfer but not under all conditions: highest transfer to secondary substrate from cotton deposit area, no apparent pattern as to where most DNA is transferred to when plastic primary substrate; loose bags increase transfer from cotton substrates; weight (68 g) does not influence transfer; for 50 µl deposits, significantly higher transfer with down placement of the bag; significantly higher transfer to primary and secondary substrate for 5µl compared to 50µl deposits on cotton

Raised Questions

possibility of a greater weight having more influence on transfer rates; is the loss of DNA during the extraction process uniform?

Cautionary Remarks

high variability between replicates (especially for plastic substrates); absolute amounts of DNA transferred/deposited not shown; simplified assumptions: extraction efficiencies equal from all substrates, transfer rates not dependent on variable initial deposit