The San Diego Conference Cool Tools and Hot Applications November 18–20, 2004 San Francisco, CA

[摘要] ### H. Akhavan-Tafti, R. DeSilva, W.G. Cripps, R.A. Eickholt, R.S. Handley, K.S. Lauwers, M.D. Sandison, S. Siripurapu, and A.P. Schaap.#### Lumigen Inc., Southfield, MI.A new type of solid phase material for binding and releasing nucleic acids has been developed. The materials bind DNA and RNA with a strength far exceeding conventional materials and feature a novel mechanism to release bound nucleic acids. Unlike silica particle based materials and ion exchange polymeric resins, both of which elute bound nucleic acid through alteration of buffer ionic strength, neither high nor low ionic strength buffer is required to release nucleic acid. Bound nucleic acid is, however, readily released intact under controlled conditions with a 5 minute protocol. The solid phase materials have been fabricated as both magnetic and non-magnetic microparticles. No chaotropic salts are required to bind or release nucleic acids. DNA and RNA can be captured from water or a variety of buffers and media over a range of pH and salt concentrations. Binding of DNA and RNA from solution results in immobilization of nucleic acids that resists reversal under all commonly used release conditions. Repeated washing of the particles with water, low or high ionic strength buffers and surfactants does not remove captured nucleic acid. Nucleic acids of a wide range of molecular weight were isolated with the new materials. DNA ranging in size from oligonucleotides up to genomic fragments of several kilobases were captured quantitatively and released in high yield. Both magnetic and non-magnetic particles have been used in methods of isolating or purifying nucleic acids from several different sources. The particles were used to isolate the plasmid pUC18 from an overnight E. coli culture after pelleting, SDS lysis and potassium acetate precipitation. Human genomic DNA was extracted from whole blood with a conventional alkaline lysis procedure. DNA in the supernatants was captured on 10 mg of particles and released to produce DNA of MW ≥ 20kb. In addition, a simple and rapid …

[学科分类] 过敏症与临床免疫学