Morphological study of fragmented DNA on touched objects.
FSI Genetics, 2008
Authors
Journal
FSI Genetics
Study Design
Addressed Question
analysis of touch DNA composition
Activity Context
Category
Specifications
Variables of Interest
Stringency of Control
Number of Individuals
2+6
Replicates per Individual and Condition
1
Nucleic Acid
Bodily Origin
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
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
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?