Nucleic acids, chromatographic methods

It is often desirable to go through a postlysis separation/concentration step prior to chromatography. Concentration methods involve the use of ammonium acetate and polyethylene glycol precipitation to further remove host proteins and small nucleic acids. These methods also reduce the volume of the sample (or the process streams) prior to chromatographic purification. The separation may also involve centrifugation and filtration to remove cell debris. 

HPLC in Nucleic Acid Research Methods and Applications, edited by Phyllis R. Brown 29. Pyrolysis and GC in Polymer Analysis, edited by S. A. Liebman andE. J. Levy 30. Modem Chromatographic Analysis of the Vitamins, edited by Andre P. De Leenheer, Willy E. 

See also Capillary Electrophoresis Clinical Applications. Forensic Sciences DNA Profiling. Lab-on-a-Chip Technologies. Mass Spectrometry Polymerase Chain Reaction Products. Nucleic Acids Chromatographic and Electrophoretic Methods. 

See also Liquid Chromatography Principles Normal Phase Reversed Phase Instrumentation. Nucleic Acids Chromatographic and Electrophoretic Methods Spectroscopic Methods. Optical Spectroscopy Stray Light. Ozone. Paints Water-Based Organic Solvent-Based. Peptides. Personal Monitoring Passive. 

See also. Chemiluminescence Oven/iew. Derivatiza-tion of Analytes. Electrophoresis Oven/iew. Enzymes Oven/iew Immobiiized Enzymes Enzyme-Based Eiec-trodes Enzymes in Physioiogicai Sampies Industriai Products and Processes Enzyme-Based Assays. Fluorescence Clinical and Drug Applications. Immunoassays Overview Production of Antibodies. Immunoassays, Applications Clinical Food Forensic. Immunoassays, Techniques Radioimmunoassays Enzyme Immunoassays Luminescence Immunoassays. Mass Spectrometry Polymerase Chain Reaction Products. Nucleic Acids Chromatographic and Electrophoretic Methods Electrochemical Methods. Polymerase Chain Reaction. 

Classical gel electrophoresis has been used extensively for protein and nucleic acid purification and characterization [9, 10], but has not been used routinely for small molecule separations, other than for polypeptides. A comparison between TLC and electrophoresis reveals that while detection is usually accomplished off-line in both electrophoretic and TLC methods, the analyte remains localized in the TLC bed and the mobile phase is immediately removed subsequent to chromatographic development. In contrast, in gel electrophoresis, the gel matrix serves primarily as an anti-... 

Mueller [103-104] has adopted the polyethylene glycol)-dextran partition system pioneered by Albertson [105] to develop an efficient chromatographic method for the resolution of proteins and nucleic acids. He used supports such... 

However, during the past three decades, an analytical method has been developed that currently rivals and may soon surpass the traditional liquid chromatographic techniques in importance for analytical separations. This technique, high-performance liquid chromatography (HPLC), is ideally suited for the separation and identification of amino acids, carbohydrates, lipids, nucleic acids, proteins, pigments, steroids, pharmaceuticals, and many other biologically active molecules. 

Highly purified DNA may be obtained by repeating the chloroform-isoamyl alcohol extraction several times. The alcohol precipitation step may also be carried out many times. Various chromatographic methods including ion exchange have been applied to the purification of nucleic acids. 

Following the discovery of DNA structure and the genetic code, various associated techniques involving the manipulation and separation of DNA fragments have been developed. Several of the physical methods are useful in nucleic-acid biochemistry these include, centrifugation, electrophoresis and chromatographic separation of DNA fragments. 

Development of an enzymic hydrolysis by Klein32 afforded the possibility of preparing with comparative ease quantities of mixed nucleotides from desoxypentose nucleic acid, but the problem remained of obtaining the individual compounds in pure form by separation of the mixture. The development of chromatographic methods and of ion-... 

Oxygen free radicals are elaborated during ischemia and reperfusion and have been strongly implicated in the pathophysiology of ischemic brain injury (114). These reactions may lead to oxidative injury to cellular lipids, proteins, and nucleic acids. Evidence for free radical elaboration in ischemia is obtainable by means of a microdialysis method in which administered salicylate is converted, in the presence of hydroxyl radical, to dihydroxybenzoic acid (DHBA) species—stable adducts detectable by chromatographic methods. A study from our laboratory of... 

As mentioned before, a number of these methods are discussed at length in this book (see Chapters 2-6). The methodologies for separation of nucleic acids, oligonucleotides, and monoclonal antibodies are covered in Chapters 8, 14, and 15. The following chromatographic methods, as they relate to separations of monoclonal antibodies, are discussed in Chapter 15 ... 

---