Cargando…

lobChIP: from cells to sequencing ready ChIP libraries in a single day

BACKGROUND: ChIP-seq is the method of choice for genome-wide studies of protein–DNA interactions. We describe a new method for ChIP-seq sample preparation, termed lobChIP, where the library reactions are performed on cross-linked ChIP fragments captured on beads. RESULTS: The lobChIP method was foun...

Descripción completa

Detalles Bibliográficos
Autores principales: Wallerman, Ola, Nord, Helena, Bysani, Madhusudhan, Borghini, Lisa, Wadelius, Claes
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4507313/
https://www.ncbi.nlm.nih.gov/pubmed/26195988
http://dx.doi.org/10.1186/s13072-015-0017-5
_version_ 1782381779126059008
author Wallerman, Ola
Nord, Helena
Bysani, Madhusudhan
Borghini, Lisa
Wadelius, Claes
author_facet Wallerman, Ola
Nord, Helena
Bysani, Madhusudhan
Borghini, Lisa
Wadelius, Claes
author_sort Wallerman, Ola
collection PubMed
description BACKGROUND: ChIP-seq is the method of choice for genome-wide studies of protein–DNA interactions. We describe a new method for ChIP-seq sample preparation, termed lobChIP, where the library reactions are performed on cross-linked ChIP fragments captured on beads. RESULTS: The lobChIP method was found both to reduce time and cost and to simplify the processing of many samples in parallel. lobChIP has an early incorporation of barcoded sequencing adaptors that minimizes the risk of sample cross-contamination and can lead to reduced amount of adaptor dimers in the sequencing libraries, while allowing for direct decross-linking and amplification of the sample. CONCLUSIONS: With results for histone modifications and transcription factors, we show that lobChIP performs equal to or better than standard protocols and that it makes it possible to go from cells to sequencing ready libraries within a single day. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13072-015-0017-5) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4507313
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-45073132015-07-21 lobChIP: from cells to sequencing ready ChIP libraries in a single day Wallerman, Ola Nord, Helena Bysani, Madhusudhan Borghini, Lisa Wadelius, Claes Epigenetics Chromatin Methodology BACKGROUND: ChIP-seq is the method of choice for genome-wide studies of protein–DNA interactions. We describe a new method for ChIP-seq sample preparation, termed lobChIP, where the library reactions are performed on cross-linked ChIP fragments captured on beads. RESULTS: The lobChIP method was found both to reduce time and cost and to simplify the processing of many samples in parallel. lobChIP has an early incorporation of barcoded sequencing adaptors that minimizes the risk of sample cross-contamination and can lead to reduced amount of adaptor dimers in the sequencing libraries, while allowing for direct decross-linking and amplification of the sample. CONCLUSIONS: With results for histone modifications and transcription factors, we show that lobChIP performs equal to or better than standard protocols and that it makes it possible to go from cells to sequencing ready libraries within a single day. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13072-015-0017-5) contains supplementary material, which is available to authorized users. BioMed Central 2015-07-21 /pmc/articles/PMC4507313/ /pubmed/26195988 http://dx.doi.org/10.1186/s13072-015-0017-5 Text en © Wallerman et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Wallerman, Ola
Nord, Helena
Bysani, Madhusudhan
Borghini, Lisa
Wadelius, Claes
lobChIP: from cells to sequencing ready ChIP libraries in a single day
title lobChIP: from cells to sequencing ready ChIP libraries in a single day
title_full lobChIP: from cells to sequencing ready ChIP libraries in a single day
title_fullStr lobChIP: from cells to sequencing ready ChIP libraries in a single day
title_full_unstemmed lobChIP: from cells to sequencing ready ChIP libraries in a single day
title_short lobChIP: from cells to sequencing ready ChIP libraries in a single day
title_sort lobchip: from cells to sequencing ready chip libraries in a single day
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4507313/
https://www.ncbi.nlm.nih.gov/pubmed/26195988
http://dx.doi.org/10.1186/s13072-015-0017-5
work_keys_str_mv AT wallermanola lobchipfromcellstosequencingreadychiplibrariesinasingleday
AT nordhelena lobchipfromcellstosequencingreadychiplibrariesinasingleday
AT bysanimadhusudhan lobchipfromcellstosequencingreadychiplibrariesinasingleday
AT borghinilisa lobchipfromcellstosequencingreadychiplibrariesinasingleday
AT wadeliusclaes lobchipfromcellstosequencingreadychiplibrariesinasingleday