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Characterization of human DNA in environmental samples.

Forensic Science International, 2008

Study Design

Addressed Question

analysis of the presence and composition of DNA from environmental dust samples

Activity Context

None

Category

Background DNARecovery

Specifications

BG on Public ItemsDNA Profiling

Variables of Interest

sampling area (human traffic level, dust visibility level)

Stringency of Control

Reality

Number of Individuals

36 samples from different areas

Replicates per Individual and Condition

1

Nucleic Acid

DNAmtDNA

Bodily Origin

trace

Depositor & Contact

Depositor Characteristics

N/A

Criteria for Shedder Status

N/A

Previous Activities

N/A

Contact Scenario

no alteration of sampling site prior to sampling - sampling

Primary Substrate

Primary Substrate Type

surfaces within academic building with low, medium or high human traffic

Primary Substrate Material

Dust

Deposit

dust visibility: unapparent, low, medium, high

Delay

N/A

Secondary Substrate

Secondary Substrate Type

N/A

Secondary Substrate Material

N/A

Secondary Substrate Contact

N/A

Further Transfer

N/A

Sampling

Background DNA on Sampled Surface

Sampled

Sampling Time

direct/delayed

Persistence

N/A

Sampling Method

sterile cotton-tipped swabs moistened with 1.7 ml cell lysis solution

Sampling Area

576 cm^2 area

Laboratory Analysis

Extraction

Mo Bio UltraClean BloodSpin purification kit, final extraction volume: 50 µl

DNA Quantification

total DNA: µQuant MQX200 plate reader, human DNA: Quantifiler Human DNA Quantification kit

Input for Profiling

1-2.5 ng or up to 20 µl of extract

Profiling

AmpFlSTR Profiler Plus PCR amplification kit, MJ BaseStation 51 DNA fragment analyzer, Cartographer v1.2.6sg and ABI Prism 3100 Avant, ABI GeneMapper ID Software v3.2, threshold: 75 rfu

Reference Samples

not available

Profile Interpretation and Mixture Analysis

allele calling following a protocol validated for casework samples and checking by two operators

RNA Data Interpretation

N/A

Results

DNA Quantity

9-28 ng/cm^2 total DNA, 0.2-1.1 pg/cm^2 human DNA

Profile Quality

partial profiles (avg. 0-5 loci/sample) generated from 25/36 samples

Parameter Used for Comparison

total DNA, human DNA (presence and quantity), Genotype recovery (= number of STR loci amplified)

Summary of Results

PCR of human nuclear TAP25 locus and mtDNA locus: human DNA present in almost all samples; high variability in amounts of DNA detected, no clear trends detectable among location, traffic level, total DNA, human DNA or percentage human to total DNA; STR recovery not based on human traffic or visible dust level but dependent on input DNA; high MW loci most often drop-out -> degradation of DNA in the sample; presence of multiple contributors detected; MJ Base Station 51 reveals more peaks/locus

Raised Questions

presence of mtDNA vs. Nuclear DNA <-> age of sample?; implementation of WGA on environmental dust samples in order to improve genotyping results

Cautionary Remarks

additional environmental variables (e.g. light, heat, moisture, cleaning protocols, ventilation systems) not taken into account