Rapid purification of double-stranded DNA by triple-helix-mediated affinity capture

Journal Article · Sat May 15 00:00:00 EDT 1993 · Analytical Chemistry (Washington); (United States)

DOI:https://doi.org/10.1021/ac00058a005· OSTI ID:6012465

Ji, H; Smith, L M ^[1]

---

1. Univ. of Wisconsin, Madison (United States)

A simple and rapid method for the preparation of highly pure plasmid DNA has been developed. The DNA is directly captured from bacterial cell lysates by formation of a triple-helical structure between the plasmid dsDNA and a 20-base biotinylated oligonucleotide attached to streptavidin-coated magnetic beads and then eluted from the beads in pH 9 buffer solution. No phenol extraction, ethanol precipitation, RNase digestion, or CsCl gradient centrifugation is required. A general purpose cloning vector, pHJ19, was constructed for this application from pUC19 DNA by insertion of a 40-base sequence suitable for triple-helix formation. The approach was also found suitable for the purification of [lambda] bacteriophage DNA. 32 refs., 6 figs., 1 tab.

• DOE Contract Number: FG02-90ER61026
• OSTI ID: 6012465
• Journal Information: Analytical Chemistry (Washington); (United States), Journal Name: Analytical Chemistry (Washington); (United States) Vol. 65:10; ISSN 0003-2700; ISSN ANCHAM
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
400100 -- Analytical & Separations Chemistry
550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
BACTERIA
BACTERIOPHAGES
CHROMOSOMES
CLONING
DNA
DNA HYBRIDIZATION
DNA-CLONING
HYBRIDIZATION
MICROORGANISMS
MOLECULAR STRUCTURE
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PARASITES
PURIFICATION
VIRUSES