Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (305K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Hagberg, A.
Right arrow Articles by Landegren, U.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hagberg, A.
Right arrow Articles by Landegren, U.
Related Collections
Right arrow Miscellaneous/other
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2000, Vol. 28, No. 11 E54-e54
© 2000 Oxford University Press

Expression profiling across many samples via manifold-assisted mRNA processing

Anette Hagberg1, Gisela Barbany3, Henrik Krook2, Martina Samiotaki4 and Ulf Landegren1,3,*

1Department of Genetics and Pathology and 2Department of Clinical Immunology, Rudbeck Laboratory, Se-75185 Uppsala, Sweden, 3Department of Medical Sciences, Uppsala University Hospital, Se-75185 Uppsala, Sweden and 4Hellenic Anticancer Institute, Papanicolaou Research Center of Oncology and Experimental Surgery, 115 22 Athens, Greece

Analysis of mRNA provides a condensed view of gene structure, and quantitative analyses can reveal induction of physiological or pathological gene expression programs. One of the main hurdles for routine mRNA analyses is the need to prepare large sets of samples in a rapid and standardized manner. We describe here a procedure for mRNA isolation and cDNA synthesis using manifold devices, consisting of a set of prongs that project into individual reaction wells. The prongs have a high binding capacity for the polyA-tails of mRNA and the captured mRNA is directly used to synthesize cDNA on the supports, followed by amplification. The convenience and reproducibility of the procedure allows profiling of gene expression over time, by comparing many different samples. Using the device mRNA was simultaneously isolated and accurately measured from up to 96 different samples of anywhere between 10 and 200 000 cells. The amounts of a leukemia-specific transcript could be measured when the malignant cells represented <=0.01% of the sampled cells. We illustrate the possibility of analyzing a number of tissues and monitoring expression of sets of cytokines, involved in rejection, at variable times after transplantation.

* To whom correspondence should be addressed at: Department of Genetics and Pathology, Rudbeck Laboratory, Se-75185 Uppsala, Sweden. Tel: +46 18 471 4910; Fax: +46 18 471 4808; Email: ulf.landegren@genpat.uu.se


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Clin. Chem.Home page
G. Barbany, A. Hagberg, U. Olsson-Stromberg, B. Simonsson, A.-C. Syvanen, and U. Landegren
Manifold-assisted Reverse Transcription-PCR with Real-Time Detection for Measurement of the BCR-ABL Fusion Transcript in Chronic Myeloid Leukemia Patients
Clin. Chem., July 1, 2000; 46(7): 913 - 920.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.