Skip Navigation

This Article
Right arrow Print PDF (2598K)
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 Search for citing articles in:
ISI Web of Science (17)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Boyd, A.C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Boyd, A.C.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1993, Vol. 21, No. 4 817-821
© 1993

METHODS

Turbo cloning: a fast, efficient method for cloning PCR products and other blunt-ended DNA fragments into plasmids

A.Christopher Boyd

MRC Human Genetics Unit, Western General Hospital Crewe Road, Edinburgh EH4 2XU, UK

Received December 24, 1992. Revised January 25, 1993. Accepted January 25, 1993.

The method uses a novel plasmid vector, p9lox5, containing a site-specific recombination sequence lox from the lox/Cre recombinase system of bacteriophage P1. There are two distinct stages. Firstly, vector and fragment DNAs are ligated intermolecularly under conditions of macromolecular crowding (15% polyethylene glycol 6000) which accelerate blunt-end Joining a thousandfold. Secondly, circular recombinant molecules are efficiently excised from the ligation products by Cre recombinase acting on pairs of lox sites within directly repeated vector molecules flanking insert DNA. Recombinants are introduced into cells conventionally by transformation or electroporation. In both a model system and the cloning of PCR products, yields approaching those obtainable in cohesive-end cloning were achieved. Applications of the technique to cDNA library generation and recovery of DNA from archive material are discussed.


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
 Infect. Immun.Home page
S.-M. Jiang, L. Wang, and P. R. Reeves
Molecular Characterization of Streptococcus pneumoniae Type 4, 6B, 8, and 18C Capsular Polysaccharide Gene Clusters
Infect. Immun., March 1, 2001; 69(3): 1244 - 1255.
[Abstract] [Full Text] [PDF]

Home page
 Genome ResHome page
J. L. Hartley, G. F. Temple, and M. A. Brasch
DNA Cloning Using In Vitro Site-Specific Recombination
Genome Res., November 1, 2000; 10(11): 1788 - 1795.
[Abstract] [Full Text]

Home page
 Hum Mol GenetHome page
J. K. Millar, J. C. Wilson-Annan, S. Anderson, S. Christie, M. S. Taylor, C. A. M. Semple, R. S. Devon, D. M. S. Clair, W. J. Muir, D. H. R. Blackwood, et al.
Disruption of two novel genes by a translocation co-segregating with schizophrenia
Hum. Mol. Genet., May 22, 2000; 9(9): 1415 - 1423.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
A. J. W. Paige, K. J. Taylor, A. Stewart, J. G. Sgouros, H. Gabra, G. C. Sellar, J. F. Smyth, D. J. Porteous, and J. E. V. Watson
A 700-kb Physical Map of a Region of 16q23.2 Homozygously Deleted in Multiple Cancers and Spanning the Common Fragile Site FRA16D
Cancer Res., March 1, 2000; 60(6): 1690 - 1697.
[Abstract] [Full Text]


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.