Automated Processing of Forensic Casework Samples Using Robotic Workstations Equipped with Nondisposable Tips: Contamination Prevention

Frégeau, Chantal J.; Lett, C. Marc; Elliott, Jim; Yensen, Craig; Fourney, Ron M.

doi:10.1111/j.1556-4029.2008.00697.x

Cited by 48 publications

(39 citation statements)

References 49 publications

(81 reference statements)

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“…The number of amplification reruns could be reduced by quantifying the extraction before STR amplification and subsequent normalization of template DNA. However, the cost of quantifying all samples using real-time PCR 8 was estimated to exceed the cost of the additional amplification reruns. Furthermore, a quantification step would prolong the overall process time with approx.…”

Section: Pcr Amplificationmentioning

confidence: 99%

Automated Extraction of DNA from Blood and PCR Setup using a Tecan Freedom EVO Liquid Handler for Forensic Genetic STR Typing of Reference Samples

Stangegaard

Frøslev

Frank-Hansen

et al. 2011

JALA: Journal of the Association for Laboratory Automation

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W e have implemented and validated automated protocols for DNA extraction and PCR setup using a Tecan Freedom EVO liquid handler mounted with the Te-MagS magnetic separation device (Tecan, M€ annedorf, Switzerland). The protocols were validated for accredited forensic genetic work according to ISO 17025 using the Qiagen MagAttract DNA Mini M48 kit (Qiagen GmbH, Hilden, Germany) from fresh whole blood and blood from deceased individuals. The workflow was simplified by returning the DNA extracts to the original tubes minimizing the risk of misplacing samples. The tubes that originally contained the samples were washed with MilliQ water before the return of the DNA extracts. The PCR was setup in 96-well microtiter plates. The methods were validated for the kits: AmpF[STR Identifiler, SGM Plus and Yfiler (Applied Biosystems, Foster City, CA), GenePrint FFFL and PowerPlex Y (Promega, Madison, WI). The automated protocols allowed for extraction and addition of PCR master mix of 96 samples within 3.5 h. In conclusion, we demonstrated that (1) DNA extraction with magnetic beads and (2) PCR setup for accredited, forensic genetic short tandem repeat typing can be implemented on a simple automated liquid handler leading to the reduction of manual work, and increased quality and throughput. ( JALA 2011;16:134-40)

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Section: Pcr Amplificationmentioning

confidence: 99%

Automated Extraction of DNA from Blood and PCR Setup using a Tecan Freedom EVO Liquid Handler for Forensic Genetic STR Typing of Reference Samples

Stangegaard

Frøslev

Frank-Hansen

et al. 2011

JALA: Journal of the Association for Laboratory Automation

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“…The second requirement for PCR is the use of synthetic oligonucleotides at least [18][19][20] nucleotides long with a defi ned sequence. Such synthetic nucleotides can be readily produced with automated instruments based on the standard reaction scheme.…”

Section: Pcrmentioning

confidence: 99%

Signal to cut-off (S/CO) ratio and detection of HCV genotype 1 by real-time PCR one-step method: is there any direct relationship?

Albertoni¹,

Arnoni²,

Araújo³

et al. 2010

Braz J Infect Dis

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Background: Polymerase chain reaction (PCR) methods play an essential role in providing data related to diagnosis, monitoring and treatment of hepatitis C virus (HCV) infection. EIA results are reported as "reactive" or "non reactive" and EIA S/CO ratio may also be reported as "high" or "low." This study aimed to evaluate the performance of a real-time RT-PCR and assess whether there is relationship between S/CO and PCR results. Study Design and Methods: Sera from blood donors were analyzed by Enzyme-Linked Immunosorbent Assay (ELISA) and RT-PCR assay to detect HCV infection. Results: The RT-PCR assay to genotypes 1a/b showed an acceptable linear response in serial dilutions. The samples were divided into two groups based on their serological results: group A -S/CO ratio < 3 (60 samples) and group B -S/CO ratio > 3 (41 samples). Viral loads were confi rmed positive in group B samples in 90%, and in group A samples were confi rmed positive in only 13% by RT-PCR. Conclusion: The methodology used was able to detect the presence of RNA-HCV genotype I in 90% of the samples serologically positive in group B. All negative samples were sent to search for other genotypes of HCV (genotypes 2-6) and were confi rmed as negative. These data suggests that these negative samples may have HCV RNA viral load below the detection limit of our test (310 IU/ mL), or a false positive result in serological test, or spontaneous viral clearance occurred.

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“…Cells were harvested using trypsin/EDTA and 1.2 × 10 4 cells/well were seeded into black 384-well clear-bottom plates (Corning Life Sciences, Lowell, MA) and incubated overnight at 37 °C, 5%CO 2 . On the day of the experiment, the media were replaced with 20 µL of calcium loading buffer and incubated for a further 45 min.…”

Section: Calcium Mobilization Assaymentioning

confidence: 99%

“…Consequently, robotic systems have found widespread utility in pharmaceutical, biotechnology, forensic science, environmental science, and clinical diagnostic laboratories. [1][2][3] Within the pharmaceutical industry, the handling and transfer of chemical compounds represents a critical step within the drug screening workflow. The precise dispensing and transfer of large numbers of compounds with diverse physicochemical properties represents an important challenge for drug discoverers.…”

Section: Introductionmentioning

confidence: 99%

The Use of Cold Plasma Technology to Reduce Carryover in Screening Assays

et al. 2013

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The accurate transfer of biological reagents represents a fundamental step in the drug screening process, and the elimination of carryover is critical for the generation of accurate measurements of biological activity. The introduction of automated liquid robotics into screening laboratories has transformed the drug screening process, enabling accurate and reproducible transfer of liquids to become a high-throughput activity, but has also introduced a new challenge for drug discoverers: to establish screening workflows that limit analyte carryover for the generation of high-quality screening data. The widespread use of pipetting tips on automated liquid handlers often necessitates the use of optimized wash protocols for removing contaminants and frequently requires the use and disposal of large quantities of organic solvents. Furthermore, many chemical and biological reagents are recalcitrant to removal from pipetting tips by treatment with organic solvents. The use of cold atmospheric plasma technology provides an alternative approach for removal of contaminants and offers many advantages over traditional decontamination protocols commonly used during biological screening. This report describes the evaluation of a cold plasma tip-cleaning system for reducing carryover in a range of biological screening assays requiring the transfer of low molecular weight compound, nucleic acid, and bacterial liquid transfers. The validation of this technology for biological screening assays is presented, and the impact of this technology for screening workflows is discussed.

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Automated Processing of Forensic Casework Samples Using Robotic Workstations Equipped with Nondisposable Tips: Contamination Prevention

Cited by 48 publications

References 49 publications

Automated Extraction of DNA from Blood and PCR Setup using a Tecan Freedom EVO Liquid Handler for Forensic Genetic STR Typing of Reference Samples

Automated Extraction of DNA from Blood and PCR Setup using a Tecan Freedom EVO Liquid Handler for Forensic Genetic STR Typing of Reference Samples

Signal to cut-off (S/CO) ratio and detection of HCV genotype 1 by real-time PCR one-step method: is there any direct relationship?

The Use of Cold Plasma Technology to Reduce Carryover in Screening Assays

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