Implementation of complete protocols in a Teflon tube

Droplet based microfluidics continues to grow as a platform for chemical and biological reactions using small quantities of fluids, however complex protocols are rarely possible in existing devices. This paper implements a new approach to merging of drops, combined with magnetic bead manipulation, f...

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Main Authors: Walsh, E, Feuerborn, A, Cook, P
Format: Conference item
Published: Society of Photo-Optical Instrumentation Engineers (SPIE) 2017
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author Walsh, E
Feuerborn, A
Cook, P
author_facet Walsh, E
Feuerborn, A
Cook, P
author_sort Walsh, E
collection OXFORD
description Droplet based microfluidics continues to grow as a platform for chemical and biological reactions using small quantities of fluids, however complex protocols are rarely possible in existing devices. This paper implements a new approach to merging of drops, combined with magnetic bead manipulation, for the creation of ligated double-stranded DNA molecule using “Gibson assembly” chemistry. DNA assembly is initially accomplished through the merging, and mixing, of five drops followed by a thermal cycle. Then, integrating this drop merging method with magnetic beads enable the implementation of amore complete protocol consisting of nine wash steps,merging of four drop, transport of selective reagents between twelve drops using magnetic particles, followed by a thermal cycle and finally the deposition of a purified drop into an Eppendorf for downstream analysis. Gel electrophoresis is used to confirm successful DNA assembly.
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spelling oxford-uuid:606f74d1-4d21-4aa8-a2ab-fdd2d209bc0c2022-03-26T17:53:25ZImplementation of complete protocols in a Teflon tubeConference itemhttp://purl.org/coar/resource_type/c_c94fuuid:606f74d1-4d21-4aa8-a2ab-fdd2d209bc0cSymplectic Elements at OxfordSociety of Photo-Optical Instrumentation Engineers (SPIE)2017Walsh, EFeuerborn, ACook, PDroplet based microfluidics continues to grow as a platform for chemical and biological reactions using small quantities of fluids, however complex protocols are rarely possible in existing devices. This paper implements a new approach to merging of drops, combined with magnetic bead manipulation, for the creation of ligated double-stranded DNA molecule using “Gibson assembly” chemistry. DNA assembly is initially accomplished through the merging, and mixing, of five drops followed by a thermal cycle. Then, integrating this drop merging method with magnetic beads enable the implementation of amore complete protocol consisting of nine wash steps,merging of four drop, transport of selective reagents between twelve drops using magnetic particles, followed by a thermal cycle and finally the deposition of a purified drop into an Eppendorf for downstream analysis. Gel electrophoresis is used to confirm successful DNA assembly.
spellingShingle Walsh, E
Feuerborn, A
Cook, P
Implementation of complete protocols in a Teflon tube
title Implementation of complete protocols in a Teflon tube
title_full Implementation of complete protocols in a Teflon tube
title_fullStr Implementation of complete protocols in a Teflon tube
title_full_unstemmed Implementation of complete protocols in a Teflon tube
title_short Implementation of complete protocols in a Teflon tube
title_sort implementation of complete protocols in a teflon tube
work_keys_str_mv AT walshe implementationofcompleteprotocolsinateflontube
AT feuerborna implementationofcompleteprotocolsinateflontube
AT cookp implementationofcompleteprotocolsinateflontube