DNA transfer during laundering may yield complete genetic profiles.
FSI Genetics, 2016
Study Design
Addressed Question
transfer of epithelial cells from vaginal secretions during washing
Activity Context
Category
Specifications
Variables of Interest
Stringency of Control
Number of Individuals
2
Replicates per Individual and Condition
1 (5)
Nucleic Acid
Bodily Origin
Depositor & Contact
Depositor Characteristics
one pre-menopausal, one post-menopausal woman, no menstruation, no sexual contact within the last 5 days
Criteria for Shedder Status
N/A
Previous Activities
N/A
Contact Scenario
wearing of 5 different pieces of underwear for 1 day each - washing with 5 pieces of clean underwear and 1 bedsheet - sampling
Primary Substrate
Primary Substrate Type
100 % cotton fabric underwear
Primary Substrate Material
Deposit
wearing for 1 day
Delay
N/A
Secondary Substrate
Secondary Substrate Type
cotton fabric underwear
Secondary Substrate Material
Secondary Substrate Contact
washing: 30°C + cold water rinse + phosphate free detergent, medium heat drying 60 min
Further Transfer
N/A
Sampling
Background DNA on Sampled Surface
Sampling Time
direct
Persistence
N/A
Sampling Method
swabbing and cutting
Sampling Area
swabbing 40 cm^2, cuttings 6.25 cm^2, 6-7 different areas per piece of underwear and sampling method
Laboratory Analysis
Extraction
differential extraction DNA IQ magnetic beads system (Promega)
DNA Quantification
Quantifiler Duo DNA Quantification kit (AB), 7500 Real-Time PCR
Input for Profiling
set volume 15 µl (min 0.008 ng/µl for epithelia fraction, 0.001 ng/µl for spermatozoa fraction)
Profiling
AmpFlSTR Identifiler Plus PCR Amplification kit 28 cycles, ABI PRISM 3130xI Genetic Analyzer, GeneMapper ID software v.3.2, AT: 50rfu, ST: 200 rfu
Reference Samples
taken from wearers
Profile Interpretation and Mixture Analysis
determination of number of alleles from reference profiles
RNA Data Interpretation
N/A
Results
DNA Quantity
<amplification threshold for swabbings, 0.2-3.2 ng for cuttings
Profile Quality
partial to complete profiles from samples with highest amplification results
Parameter Used for Comparison
DNA quantity, profile completeness
Summary of Results
higher amounts of DNA from pre-menopausal woman; partial to complete genetic profiles obtainable
Raised Questions
N/A
Cautionary Remarks
sample size too low for differentiation between pre- and post-menopausal stage (n=1); quality of DNA profiles (peak height contributions, unknown alleles n.a.)