Published online 2 January 2004
Nucleic Acids Research, 2004, Vol. 32, No. 1 e2
© 2004 Oxford University Press
Ligase-mediated construction of branched DNA strands: a novel DNA joining activity catalyzed by T4 DNA ligase
1 Rudbeck Laboratory, Department of Genetics and Pathology, Se-75185 Uppsala, Sweden and 2 Amersham Biosciences, Uppsala, Sweden
*To whom correspondence should be addressed. Tel: +46 18 471 4910; Fax: +46 18 471 4808; Email: ulf.landegren{at}genpat.uu.se
Present address:
Anil Kumar, Solexa Limited, Chesterford Research Park, Little Chesterford, Nr Saffron Walden, Essex CB10 1XL, UK
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
Branched nucleic acid strands exist as intermediates in certain biological reactions, and bifurcating DNA also presents interesting opportunities in biotechnological applications. We describe here how T4 DNA ligase can be used for efficient construction of DNA molecules having one 5' end but two distinct 3' ends that extend from the 2' and 3' carbons, respectively, of an internal nucleotide. The nature of the reaction products is investigated, and optimal reaction conditions are reported for the construction of branched oligonucleotides. We discuss the utility of these branched DNA nanostructures for gene detection.