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Improving recovery and stability of touch DNA

FSI Genetics Supplement Series, 2017

Study Design

Addressed Question

yield and stability of samples collected with detergent-based wetting agents in comparison to water

Activity Context

None

Category

PersistenceRecovery

Specifications

Persistence with TemperaturePersistence with TimeSamplingSurface

Variables of Interest

primary substratewetting agent

Stringency of Control

Controlled

Number of Individuals

N/A

Replicates per Individual and Condition

3

Nucleic Acid

DNA

Bodily Origin

skin (hands)

Depositor & Contact

Depositor Characteristics

N/A

Criteria for Shedder Status

N/A

Previous Activities

handwashing 5 min before handling

Contact Scenario

handwashing - handling objects - sampling - delay - extraction

Primary Substrate

Primary Substrate Type

plastic, metal and glass bottles

Primary Substrate Material

GlassMetalPlastic

Deposit

medium pressure contact for 30 s

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

Negative (Assumed)

Sampling Time

direct

Persistence

time: 24 h storage at temperature: -20°C, RT, 37°C, 50°C between sampling and extraction

Sampling Method

double swab technique using distilled water or inhouse 1% n-lauroylsarcosine buffer

Sampling Area

handled areas (clearly visible)

Laboratory Analysis

Extraction

PureGene extraction kit (Qiagen)

DNA Quantification

Quantifiler Human DNA quantification kit

Input for Profiling

1 µl

Profiling

in house assay for the amplification of four amplicons (50-154 bp), ABI 3500 Prism Genetic Analyzer; GeneMapper ID-X software v. 1.2.

Reference Samples

n.a. due to inhouse amplification assay

Profile Interpretation and Mixture Analysis

N/A

RNA Data Interpretation

N/A

Results

DNA Quantity

0-> 5ng/µl (extraction volume n.s.)

Profile Quality

full profiles, varying peak heights

Parameter Used for Comparison

DNA concentration (ng/µl), recovery peak heights

Summary of Results

The use of lysis buffer gives an overall higher DNA concentration directly after sampling in comparison to water; For storage before extraction, there was a temperature dependent deterioration of DNA with a higher stability for samples collected with detergent-based wetting agent; Conclusion: detergent-based moistening agents are more efficient for sampling and lead to an increased stability during storage than water

Raised Questions

N/A

Cautionary Remarks

details missing on experimental set-up: swab type, individual characteristics, which individuals handled which items, extraction volume; differences between materials not assessed; number of data points in Fig. 1 higher than described in methods section; surprisingly high concentrations for touch DNA (kit: 50-250 µl elution volume, thus recovery of >50 ng consistently shown here)