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Maximization of STR DNA typing success for touched objects.

International Congress Series, 2006

Study Design

Addressed Question

evaluation and comparison of different sampling, extraction and amplification methods for LT-DNA

Activity Context

None

Category

Recovery

Specifications

DNA ProfilingExtractionSampling

Variables of Interest

DNA Sourcemethods (sampling, extraction, amplification)

Stringency of Control

Controlled

Number of Individuals

N/A

Replicates per Individual and Condition

3

Nucleic Acid

DNA

Bodily Origin

HEK cellsskin (fingertips)

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

Negative (Assumed)

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