Directed assembly of DNA molecules via simultaneous ligation and digestion.

Directed assembly of DNA molecules via simultaneous ligation and digestion. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

Directed assembly of DNA molecules via simultaneous ligation and digestion. Abstract Text:

Gregory J Cost,Nicholas R Cozzarelli,

DNA ligation is a routine laboratory practice, yet the yield of the desired product is often very low due to competing off-pathway reactions. The sensitivity of subsequent manipulations (e.g., selection via bacterial transformation) often obviates the need for a high yield of correctly ligated products. However the ability to perform high-yield, preparative-scale DNA ligations would benefit a number of downstream applications ranging from standard molecular cloning to biophysics and DNA computing. We describe here a ligation technique that specifically converts off-pathway ligation products back into substrate. We term this second-chance strategy enzymatic ligation assisted by nucleases (ELAN) and demonstrate the ordered assembly of four DNA fragments via simultaneous ligation and digestion in the presence of eight restriction enzymes. Use of ELAN increased the yield of the desired product by more than 30-fold.

Directed assembly of DNA molecules via simultaneous ligation and digestion. Publishing Authors By Initials

GJ Cost,NR Cozzarelli,

For similar investigative techniques: genetic techniques research abstracts see: investigative techniques: genetic techniques research

PUBMED ID PMID:

MEDLINE DATE:

Directed assembly of DNA molecules via simultaneous ligation and digestion. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: BioTechniques

VOLUME: 42

Page Numbers: 84, 86-9

Journal Abbreviation: BioTechniques

ISSN: 0736-6205

DAY: 3

MONTH: Jan

YEAR: 2007

Directed assembly of DNA molecules via simultaneous ligation and digestion. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 8306785

Directed assembly of DNA molecules via simultaneous ligation and digestion. Keywords Mesh Terms:

KEYWORDS: Genetic Techniques

MESH TERMS: chemical synthesis

Chemical & Substance for Abstract: Directed assembly of DNA molecules via simultaneous ligation and digestion. Information

Substance Name: DNA Restriction Enzymes

Registry Number: EC 3.1.21.-

Grant and Affiliation Information for Directed assembly of DNA molecules via simultaneous ligation and digestion.

AFFILIATION: Department of Molecular and Cell Biology, University of California-Berkeley, Berkeley, CA 94720, USA. gregcost@berkeley.edu

Country: United States

AGENCY: United States NIGMS

GRANT: GM31657

ACRONYM: GM

MEDLINETA: Biotechniques

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

Directed assembly of DNA molecules via simultaneous ligation and digestion Related Publications

• Biodegradation in contaminated aquifers: incorporating microbial/molecular methods.
• Directed assembly of DNA molecules via simultaneous ligation and digestion.
• Identification of the FtsK sequence-recognition domain.
• Making back issues of PNAS online free.
• Multiple modes of Escherichia coli DNA gyrase activity revealed by force and torque.
• PNAS early edition.
• Redesign of PNAS: getting ready for the millennium.
• Redesign of PNAS: new covers for the journal.