Improving recovery and stability of touch DNA
FSI Genetics Supplement Series, 2017
Authors
Journal
FSI Genetics Supplement Series
Study Design
Addressed Question
yield and stability of samples collected with detergent-based wetting agents in comparison to water
Activity Context
Category
Specifications
Variables of Interest
Stringency of Control
Number of Individuals
N/A
Replicates per Individual and Condition
3
Nucleic Acid
Bodily Origin
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
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
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)