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A cDNA RDA protocol using solid-phase technology suited for analysis in small tissue samples.
KTH, Superseded Departments, Biotechnology.ORCID iD: 0000-0003-0996-1644
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2000 (English)In: Biomolecular Engineering, ISSN 1389-0344, E-ISSN 1878-559X, Vol. 17, no 1, 1-9 p.Article in journal (Refereed) Published
Abstract [en]

cDNA representational difference analysis (cDNA RDA) is a PCR-based subtractive enrichment procedure for the cloning of differentially expressed genes. In this study, we have further developed the procedure to take advantage of solid-phase technology, and to facilitate the use of RDA when starting material is limited. Several parameters of the PCR-based generation of cDNA representations were investigated, and a solid-phase based purification step was introduced to simplify removal of digested adapter-ends and uncleaved fragments. The use of magnetic particles increased the speed of the method, and also eliminated the risk of carry-over contamination between iterative steps of subtraction and PCR amplification. The modified protocol was evaluated in monitoring differences in gene expression in (i) a rat system consisting of livers with and without growth hormone treatment, and in (ii) a human system consisting of normal colon and colon cancer.

Place, publisher, year, edition, pages
2000. Vol. 17, no 1, 1-9 p.
National Category
Industrial Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-13363PubMedID: 11042471OAI: oai:DiVA.org:kth-13363DiVA: diva2:324420
Note
QC 20100615Available from: 2010-06-15 Created: 2010-06-15 Last updated: 2017-12-12Bibliographically approved
In thesis
1. Analysis of human gene transcripts: Identification and characterisation based on homology and differential expression
Open this publication in new window or tab >>Analysis of human gene transcripts: Identification and characterisation based on homology and differential expression
1998 (English)Doctoral thesis, comprehensive summary (Other scientific)
Place, publisher, year, edition, pages
Stockholm: KTH, 1998. 68 p.
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-2751 (URN)91-7170-347-0 (ISBN)
Public defence
1998-12-18, 00:00
Note
QC 20100615Available from: 2000-01-01 Created: 2000-01-01 Last updated: 2010-06-17Bibliographically approved

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CiteExportLink to record
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  • apa
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