Lysis of a Single Cyanobacterium for Whole Genome Amplification
Bacterial species from natural environments, exhibiting a great degree of genetic diversity that has yet to be characterized, pose a specific challenge to whole genome amplification (WGA) from single cells. A major challenge is establishing an effective, compatible, and controlled lysis protocol. We...
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doaj-3052c5ebb7234e24868ff7f647e5d5902020-11-25T00:41:55ZengMDPI AGMicromachines2072-666X2013-08-014332133210.3390/mi4030321Lysis of a Single Cyanobacterium for Whole Genome AmplificationRichard N. ZareEric W. HallVisham AppadooSamuel KimBacterial species from natural environments, exhibiting a great degree of genetic diversity that has yet to be characterized, pose a specific challenge to whole genome amplification (WGA) from single cells. A major challenge is establishing an effective, compatible, and controlled lysis protocol. We present a novel lysis protocol that can be used to extract genomic information from a single cyanobacterium of Synechocystis sp. PCC 6803 known to have multilayer cell wall structures that resist conventional lysis methods. Simple but effective strategies for releasing genomic DNA from captured cells while retaining cellular identities for single-cell analysis are presented. Successful sequencing of genetic elements from single-cell amplicons prepared by multiple displacement amplification (MDA) is demonstrated for selected genes (15 loci nearly equally spaced throughout the main chromosome).http://www.mdpi.com/2072-666X/4/3/321cyanobacterialysiswhole genome amplificationsingle cellmicrofluidics |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Richard N. Zare Eric W. Hall Visham Appadoo Samuel Kim |
spellingShingle |
Richard N. Zare Eric W. Hall Visham Appadoo Samuel Kim Lysis of a Single Cyanobacterium for Whole Genome Amplification Micromachines cyanobacteria lysis whole genome amplification single cell microfluidics |
author_facet |
Richard N. Zare Eric W. Hall Visham Appadoo Samuel Kim |
author_sort |
Richard N. Zare |
title |
Lysis of a Single Cyanobacterium for Whole Genome Amplification |
title_short |
Lysis of a Single Cyanobacterium for Whole Genome Amplification |
title_full |
Lysis of a Single Cyanobacterium for Whole Genome Amplification |
title_fullStr |
Lysis of a Single Cyanobacterium for Whole Genome Amplification |
title_full_unstemmed |
Lysis of a Single Cyanobacterium for Whole Genome Amplification |
title_sort |
lysis of a single cyanobacterium for whole genome amplification |
publisher |
MDPI AG |
series |
Micromachines |
issn |
2072-666X |
publishDate |
2013-08-01 |
description |
Bacterial species from natural environments, exhibiting a great degree of genetic diversity that has yet to be characterized, pose a specific challenge to whole genome amplification (WGA) from single cells. A major challenge is establishing an effective, compatible, and controlled lysis protocol. We present a novel lysis protocol that can be used to extract genomic information from a single cyanobacterium of Synechocystis sp. PCC 6803 known to have multilayer cell wall structures that resist conventional lysis methods. Simple but effective strategies for releasing genomic DNA from captured cells while retaining cellular identities for single-cell analysis are presented. Successful sequencing of genetic elements from single-cell amplicons prepared by multiple displacement amplification (MDA) is demonstrated for selected genes (15 loci nearly equally spaced throughout the main chromosome). |
topic |
cyanobacteria lysis whole genome amplification single cell microfluidics |
url |
http://www.mdpi.com/2072-666X/4/3/321 |
work_keys_str_mv |
AT richardnzare lysisofasinglecyanobacteriumforwholegenomeamplification AT ericwhall lysisofasinglecyanobacteriumforwholegenomeamplification AT vishamappadoo lysisofasinglecyanobacteriumforwholegenomeamplification AT samuelkim lysisofasinglecyanobacteriumforwholegenomeamplification |
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1725284871553679360 |