Polynucleotides, chromatography

Redissolve DNA in 50 pi of TE and remove ATP by spin chromatography (polynucleotides) or polyacrylamide gels (oligomers Section 6.3.2). 

It would be easier to describe those classes of compounds not normally separated by RPLC than to catalogue the applications to which RPLC has been turned. Applications for reversed phase can be found in virtually every area of analysis and are reviewed regularly in the journal Analytical Chemistry. RPLC has not been in general use for the analysis of inorganic ions, which are readily separated by ion exchange chromatography polysaccharides, which tend to be too hydrophilic to separate by RPLC polynucleotides, which tend to adsorb irreversibly to the reversed phase packing and compounds which are so hydrophobic that reversed phase offers little selectivity. 

Murphy, R.E. (2001). Comprehensive two-dimensional liquid chromatography for comparative protein analysis. Presented at the International Symposium on the Separation of Proteins, Peptides and Polynucleotides. (ISPPP 2001) Orlando, FL. 

Other methods that are related to affinity chromatography include hydrophobic interaction chromatography and thiophilic adsorption. The former is based on the interactions of proteins, peptides, and nucleic acids with short nonpolar chains on a support. This was first described in 1972 [113,114] following work that examined the role of spacer arms on the nonspecific adsorption of affinity columns [114]. Thiophilic adsorption, also known as covalent or chemisorption chromatography, makes use of immobilized thiol groups for solute retention [115]. Applications of this method include the analysis of sulfhydryl-containing peptides or proteins and mercurated polynucleotides [116]. 

DHPLC (C2, L3, U1) uses a proprietary matched ion polynucleotide chromatography column devised by Transgenomics, Inc., in combination with an organic solvent and thermal denaturation (rather than an electrophoretic gel) to resolve heteroduplexes containing a suspected polymorphism from a polymorphism-free homoduplex. Homo- and heteroduplexes are visualized through UV absorbance readings of the eluent as it leaves the column. 

McMaster, R., Kruk, J., Christianson, G., Gomez, P., Warner, T., Demmer, W., and Nussbaumer, D. (1998). Purification of clinical vaccine proteins by high performance membrane chromatography. Proceedings of 18th Int. Symp. on Sep. and Anal, of Prot., Pep. and Polynucleotides (ISPPP 98), p. 29, abstract 114, Vienna. 

Following the first studies of J.J. Thomson (1912), mass spectrometry has undergone countless improvements. Since 1958, gas chromatography coupled with mass spectrometry has revolutionized the analysis of volatile compounds. Another revolution occurred in the 1980s when the technique became available for the study of non-volatile compounds such as peptides, oligosaccharides, phospholipids, bile salts, etc. From the discoveries of electrospray and matrix-assisted laser desorption in the late 1980s, compounds with molecular masses exceeding several hundred thousands of daltons, such as synthetic polymers, proteins, gly-cans and polynucleotides, have been analysed by mass spectrometry. 

Baba Y, Matsuura T, Wakamoto K, Tsuhako M (1991a) Comparison of high-performance liquid chromatography with capillary gel electrophoresis in single base resolution of polynucleotides. J Chromatgr 558 273-284. 

Most chemical classes have been separated and analyzed by CEC [6]. These include many classes of pharmaceuticals, environmental chemicals, explosives, natural products, drugs of abuse, polypeptides, oligosaccharides, nucleosides, and their bases and polynucleotides. Applications of CEC are readily found in Analytical Abstracts for example, a publication of the American Chemical Society, or the indexes of journals such as the Journal of Chromatography. 

When the kinetics of mass transfer is slow e.g., in some applications of liqmd-solid chromatography, ion-exchange chromatography or affinity chromatography to the separation of large peptides, proteins, or polynucleotides), a relationship of the following form is written to take this into accoxmt. 

The papers published during the past year in the field of nucleotide and polynucleotide chemistry have been less remarkable for innovative chemistry than for biochemical application, and sheer volume has necessitated much pruning of the material available. The appearance of a new journal - Nucleic Acid Research - is symptomatic of the increasing publication in this area. Cyclic AMP research, Sutherland s monument, has yielded many new compounds, and no attempt has been made to cover the huge quantity of biochemical and pharmacological data available on these, for which the reader is advised to seek specific reviews. Affinity labelling and affinity chromatography continue to justify the wide research effort they command. 

---