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Whole genome amplification of labeled viable single cells suited for array-comparative genomic hybridization

Författare och institution:
Thomas Kroneis (Sahlgrenska Cancer Center); A. El-Heliebi (-)
Publicerad i:
Whole Genome Amplification: Methods and Protocols, s. 233-243
ISBN:
978-149392990-0
Publikationstyp:
Kapitel
Förlag:
Springer
Förlagsort:
New York
Publiceringsår:
2015
Språk:
engelska
Fulltextlänk:
Sammanfattning (abstract):
Understanding details of a complex biological system makes it necessary to dismantle it down to its components. Immunostaining techniques allow identification of several distinct cell types thereby giving an inside view of intercellular heterogeneity. Often staining reveals that the most remarkable cells are the rarest. To further characterize the target cells on a molecular level, single cell techniques are necessary. Here, we describe the immunostaining, micromanipulation, and whole genome amplification of single cells for the purpose of genomic characterization. First, we exemplify the preparation of cell suspensions from cultured cells as well as the isolation of peripheral mononucleated cells from blood. The target cell population is then subjected to immunostaining. After cytocentrifugation target cells are isolated by micromanipulation and forwarded to whole genome amplification. For whole genome amplification, we use GenomePlex® technology allowing downstream genomic analysis such as array-comparative genomic hybridization. © Springer Science+Business Media New York 2015. All rights reserved.
Ämne (baseras på Högskoleverkets indelning av forskningsämnen):
MEDICIN OCH HÄLSOVETENSKAP ->
Medicinska grundvetenskaper ->
Medicinsk genetik
Nyckelord:
Immunostaining, Micromanipulation, Single-cell analysis, Whole genome amplification
Postens nummer:
240419
Posten skapad:
2016-08-18 15:41
Posten ändrad:
2016-08-18 15:42

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