Development of an automated AmpliSeq™ library building workflow for biological stain samples on the Biomek® 3000

Here, we present the development of an automated AmpliSeq™ (ThermoFischer, MA, USA) workflow for library building using the Biomek® 3000 Laboratory Automation Workstation (Beckman Coulter Inc., CA, USA), in which the total volume of PCR reagents and reagents for library preparation are reduced by on...

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Main Authors: Maryam Sharafi Farzad, Brian Møllegaard Pedersen, Helle Smidt Mogensen, Claus Børsting
Format: Article
Language:English
Published: Taylor & Francis Group 2020-06-01
Series:BioTechniques
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Online Access:https://www.future-science.com/doi/10.2144/btn-2019-0156
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author Maryam Sharafi Farzad
Brian Møllegaard Pedersen
Helle Smidt Mogensen
Claus Børsting
author_facet Maryam Sharafi Farzad
Brian Møllegaard Pedersen
Helle Smidt Mogensen
Claus Børsting
author_sort Maryam Sharafi Farzad
collection DOAJ
description Here, we present the development of an automated AmpliSeq™ (ThermoFischer, MA, USA) workflow for library building using the Biomek® 3000 Laboratory Automation Workstation (Beckman Coulter Inc., CA, USA), in which the total volume of PCR reagents and reagents for library preparation are reduced by one-half. The automated AmpliSeq workflow was tested using 43 stain samples (blood, bone, muscle tissue, semen, swab, nail scrape and cigarette butts) collected from crime scenes. The sequencing data were evaluated for locus balance, heterozygous allele balance and noise. The performance of libraries built with the automated AmpliSeq workflow using one-half of the recommended reagent volumes were similar to the performance of libraries built with the recommended (full) volumes of the reagents.
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1940-9818
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publishDate 2020-06-01
publisher Taylor & Francis Group
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series BioTechniques
spelling doaj-art-822083c3c7cb4b0bbe70c2e059d574b22025-08-20T02:26:05ZengTaylor & Francis GroupBioTechniques0736-62051940-98182020-06-0168634234410.2144/btn-2019-0156Development of an automated AmpliSeq™ library building workflow for biological stain samples on the Biomek® 3000Maryam Sharafi Farzad0Brian Møllegaard Pedersen1Helle Smidt Mogensen2Claus Børsting31Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health & Medical Sciences, University of Copenhagen, Copenhagen, Denmark1Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health & Medical Sciences, University of Copenhagen, Copenhagen, Denmark1Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health & Medical Sciences, University of Copenhagen, Copenhagen, Denmark1Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health & Medical Sciences, University of Copenhagen, Copenhagen, DenmarkHere, we present the development of an automated AmpliSeq™ (ThermoFischer, MA, USA) workflow for library building using the Biomek® 3000 Laboratory Automation Workstation (Beckman Coulter Inc., CA, USA), in which the total volume of PCR reagents and reagents for library preparation are reduced by one-half. The automated AmpliSeq workflow was tested using 43 stain samples (blood, bone, muscle tissue, semen, swab, nail scrape and cigarette butts) collected from crime scenes. The sequencing data were evaluated for locus balance, heterozygous allele balance and noise. The performance of libraries built with the automated AmpliSeq workflow using one-half of the recommended reagent volumes were similar to the performance of libraries built with the recommended (full) volumes of the reagents.https://www.future-science.com/doi/10.2144/btn-2019-0156AmpliSeq™ workflowBiomek® 3000library buildingmassively parallel sequencing
spellingShingle Maryam Sharafi Farzad
Brian Møllegaard Pedersen
Helle Smidt Mogensen
Claus Børsting
Development of an automated AmpliSeq™ library building workflow for biological stain samples on the Biomek® 3000
BioTechniques
AmpliSeq™ workflow
Biomek® 3000
library building
massively parallel sequencing
title Development of an automated AmpliSeq™ library building workflow for biological stain samples on the Biomek® 3000
title_full Development of an automated AmpliSeq™ library building workflow for biological stain samples on the Biomek® 3000
title_fullStr Development of an automated AmpliSeq™ library building workflow for biological stain samples on the Biomek® 3000
title_full_unstemmed Development of an automated AmpliSeq™ library building workflow for biological stain samples on the Biomek® 3000
title_short Development of an automated AmpliSeq™ library building workflow for biological stain samples on the Biomek® 3000
title_sort development of an automated ampliseq™ library building workflow for biological stain samples on the biomek r 3000
topic AmpliSeq™ workflow
Biomek® 3000
library building
massively parallel sequencing
url https://www.future-science.com/doi/10.2144/btn-2019-0156
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