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Low-volume on-chip single-cell whole genome amplification for multiple subsequent analyses

Författare och institution:
Thomas Kroneis (Sahlgrenska Cancer Center); S. Chen (-); A. El-Heliebi (-)
Publicerad i:
Whole Genome Amplification: Methods and Protocols, s. 245-261
ISBN:
978-149392990-0
Publikationstyp:
Kapitel
Förlag:
Springer
Förlagsort:
New York
Publiceringsår:
2015
Språk:
engelska
Fulltextlänk:
Sammanfattning (abstract):
Multiple analyses such as DNA profiling, sequencing, or comparative genome hybridization (CGH) done on the single-cell level long for pre-amplification due to the diploid human genome. Isothermal whole genome amplification allows amplification of long DNA templates from single cells. When analysis needs to be performed under rare cell conditions additional care needs to be taken due to the fact that, even after pre-enrichment, few candidate target cells are still dispersed among an overwhelming number of non-target background cells. Here, we describe a protocol where we define a population of candidate target cells based on specific staining. Candidate cells are then isolated by laser microdissection and pressure catapulting (LMPC) and transferred onto a microliter reaction slide. This slide allows monitoring the single-cell isolation process and isothermal whole genome amplification in less than 2 μL. The amplification products obtained from single cells can be forwarded to multiple analyses. © 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:
AmpliGrid, Isothermal whole genome amplification, Laser microdissection, Single-cell analysis
Postens nummer:
240417
Posten skapad:
2016-08-18 15:31
Posten ändrad:
2016-08-18 15:32

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