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Morphological study of fragmented DNA on touched objects.

FSI Genetics, 2008

Study Design

Addressed Question

analysis of touch DNA composition

Activity Context

None

Category

Background DNA

Specifications

BG on Skin / Other Body LocationsBodily Origin

Variables of Interest

Sampling Methodanalysis method

Stringency of Control

Reality

Number of Individuals

2+6

Replicates per Individual and Condition

1

Nucleic Acid

DNA

Bodily Origin

skin (neck)

Depositor & Contact

Depositor Characteristics

N/A

Criteria for Shedder Status

N/A

Previous Activities

cleaning sampling area (95% ethanol) and sitting in clean room at 30°C for 30 min

Contact Scenario

cleaning sampling area - sitting at 30°C for 30 min - sampling

Primary Substrate

Primary Substrate Type

body part: neck

Primary Substrate Material

Skin

Deposit

regular presence, sweating

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

Persistence

N/A

Sampling Method

swabbing (wet: sterile water), tapelifting

Sampling Area

neck area

Laboratory Analysis

Extraction

proteolytic digestion, phenol-chloroform extraction, ethanol precipitation, final volume: 10 µl

DNA Quantification

human specific ds DNA RT PCR assay human genome quantification kit

Input for Profiling

N/A

Profiling

Ampfl STR Identifiler Kit, AB 3130 xl genetic analyzer, GeneMapper analysis software ver.3.2, threshold: 150 rfu

Reference Samples

buccal swabs taken from all individuals

Profile Interpretation and Mixture Analysis

comparison to reference profiles

RNA Data Interpretation

N/A

Results

DNA Quantity

3.0-33.3 ng

Profile Quality

full profiles

Parameter Used for Comparison

Immunostaining and EM results, presence of human DNA (qPCR and STR typing)

Summary of Results

human skin autopsies: EM shows condensed and shapeless nuclei in the upper cell layer, immunohistochemistry shows fragmented DNA in the nucleus-like objects on the skin surface and in the uppermost cornified skin layer; human skin swabbings: Immunomes showing ssDNA present in skin swabbings; agarose gel electrophorese: high MW and degraded DNA present in swabbings and tapelifts -> more degraded DNA in swabbings; STR profiles from neck skin swabs possible; conclusion: fragmented DNA constantly sloughed off cornified layers

Raised Questions

role of sweat?

Cautionary Remarks

presence of fragmented ssDNA and presence of human DNA demonstrated, no confirmation that fragmented ssDNA is human (as cells are also present), no indication whether STR profiles are derived from DNA fragments; role of sweat only hypothesized; comparability between neck and hand skin samples?