Maximization of STR DNA typing success for touched objects.
International Congress Series, 2006
Authors
- HBHoward Baum
- JSJames A. Sebestyen
- JTJeannie Tamariz
- TCTheresa Caragine
- MPMechthild Prinz
- EBEwelina Bajda
- RSRobert C. Shaler
- LSLinnea Schiffner
Journal
International Congress Series
Study Design
Addressed Question
evaluation and comparison of different sampling, extraction and amplification methods for LT-DNA
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
N/A
Contact Scenario
touch deposit or direct deposit of cells - sampling
Primary Substrate
Primary Substrate Type
flat surface
Primary Substrate Material
N/A
Deposit
fingerprint deposit, n.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
N/A
Sampling Method
swabbing (cotton, Dacron or "specialized swabs") moistened with 0.01% SDS
Sampling Area
deposit areas
Laboratory Analysis
Extraction
SDS and proteinase K, Microcon 100 column
DNA Quantification
qPCR on Rotorgene 3000
Input for Profiling
6.25-1000 pg
Profiling
AmpFlSTR Identifiler or Powerplex 16, 31 or 32 cycles (LCN), 3100 Genetic Prism Analyzer, threshold: 75 rfu, only alleles appearing in min. 2/3 amplifications assigned
Reference Samples
N/A
Profile Interpretation and Mixture Analysis
assignment of alleles occurring in 2/3 amplifications
RNA Data Interpretation
N/A
Results
DNA Quantity
6.25-1000 pg
Profile Quality
full profiles from 25 pg input and more
Parameter Used for Comparison
profile quality (clean codis, partial, mixture, inconclusive)
Summary of Results
"specialized swabs" more effective than common swabbing material; SDS digestion + Microcon concentration more effective than "protocols with many manipulations"; most effective profiling method: Identifiler kit with doubled annealing time, increased cycle number and reduced reaction volume and amplification in triplicates;
Raised Questions
N/A
Cautionary Remarks
data mostly not shown; several details on touched samples and methods missing