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

Kapitel i bok
Författare Thomas Kroneis
A. El-Heliebi
Publicerad i Whole Genome Amplification: Methods and Protocols
Sidor 233-243
ISBN 978-149392990-0
Förlag Springer
Förlagsort New York
Publiceringsår 2015
Publicerad vid Sahlgrenska Cancer Center
Sidor 233-243
Språk en
Länkar dx.doi.org/10.1007/978-1-4939-2990-...
Ämnesord Immunostaining, Micromanipulation, Single-cell analysis, Whole genome amplification
Ämneskategorier Medicinsk genetik

Sammanfattning

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.

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