Skip Navigation


Nucleic Acids Research Advance Access first published online on February 22, 2007
This version published online on March 13, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm078
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (138K) Freely available
Right arrow Screen PDF (228K) Freely available
Right arrow Supplementary Material
Right arrowOA All Versions of this Article:
35/7/e47    most recent
gkm078v2
gkm078v1
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 arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Fredriksson, S.
Right arrow Articles by Davis, R. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fredriksson, S.
Right arrow Articles by Davis, R. W.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Multiplex amplification of all coding sequences within 10 cancer genes by Gene-Collector

Simon Fredriksson*, Johan Banér, Fredrik Dahl, Angela Chu, Hanlee Ji, Katrina Welch and Ronald W. Davis

Stanford Genome Technology Center, Bio-X, 318 Campus Dr., Stanford, California 94305, USA

*To whom correspondence should be addressed. Tel: +1-650-723-6287; Fax: +1-650-724-5791; Email: simon.fredriksson{at}stanford.edu

Received December 29, 2006. Revised January 23, 2007. Accepted January 24, 2007.

Herein we present Gene-Collector, a method for multiplex amplification of nucleic acids. The procedure has been employed to successfully amplify the coding sequence of 10 human cancer genes in one assay with uniform abundance of the final products. Amplification is initiated by a multiplex PCR in this case with 170 primer pairs. Each PCR product is then specifically circularized by ligation on a Collector probe capable of juxtapositioning only the perfectly matched cognate primer pairs. Any amplification artifacts typically associated with multiplex PCR derived from the use of many primer pairs such as false amplicons, primer-dimers etc. are not circularized and degraded by exonuclease treatment. Circular DNA molecules are then further enriched by randomly primed rolling circle replication. Amplification was successful for 90% of the targeted amplicons as seen by hybridization to a custom resequencing DNA micro-array. Real-time quantitative PCR revealed that 96% of the amplification products were all within 4-fold of the average abundance. Gene-Collector has utility for numerous applications such as high throughput resequencing, SNP analyses, and pathogen detection.


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
 Cancer Res.Home page
K. H. Taylor, R. S. Kramer, J. W. Davis, J. Guo, D. J. Duff, D. Xu, C. W. Caldwell, and H. Shi
Ultradeep Bisulfite Sequencing Analysis of DNA Methylation Patterns in Multiple Gene Promoters by 454 Sequencing
Cancer Res., September 15, 2007; 67(18): 8511 - 8518.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
F. Dahl, J. Stenberg, S. Fredriksson, K. Welch, M. Zhang, M. Nilsson, D. Bicknell, W. F. Bodmer, R. W. Davis, and H. Ji
Multigene amplification and massively parallel sequencing for cancer mutation discovery
PNAS, May 29, 2007; 104(22): 9387 - 9392.
[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.