Nucleic acid techniques electrophoresis

R. C. Allen and B. Budowle. Gel electrophoresis of proteins and nucleic acids selected techniques. Walter de Gruyter, Berlin (1994). 

The refinement of other analytical methods, such as electrophoresis [34,36], the various techniques of optical spectroscopy [103-105], and nuclear magnetic resonance [201], is supplemented by the recent advances in real-time affinity measurements [152,202], contributing to the understanding of biomolecular reactivity. Taken together, the improvement of analytical methods will eventually allow a comprehensive characterization of the structure, topology, and properties of the nucleic acid-based supramolecular components under consideration for distinctive applications in nanobiotechnology. 

Diamandis, E.P. (1993) Time-resolved fluorometry in nucleic acid hybridization and Western blotting techniques (Review). Electrophoresis 14, 866-875. 

Biomolecular MS and in particular MALDI-TOF-MS (see Sections 2.1.22 and 2.2.1) permit the routine analysis of oligonucleotides up to 70-mers, intact nucleic acids, and the direct detection of DNA products with no primer labels with an increase in analysis speed and mass accuracy especially in contrast to traditional DNA separation techniques such as slab gels or capillary electrophoresis. Applications focus on the characterization of single nucleotide polymorphisms (SNPs) and short tandem repeats (STRs). Precise and accurate gene expression measurements show relative and absolute numbers of target molecules determined independently of the number of PCR cycles. DNA methylation can be studied quantitatively. 

Electrophoresis has been applied for decades in analytical chemistry. It is still the method of choice for the determination of proteins and nucleic acids. The technique of CE is classified in three main groups of methods [1] ... 

As discussed in Chapter 11, electrophoresis refers to a group of techniques used to separate and study molecules with electrical charges. Based upon these charges, both sign and magnitude, biomolecules such as proteins, peptides, amino acids, nucleic acids, and fragmented nucleic acids migrate in an electrical... 

Electrophoresis is normally run in aqueous media, hence the analytes must be soluble in water. Presently only three types of water-soluble dendrimers have been successfully analyzed using gel electrophoresis techniques. The list includes Starburst PAMAM dendrimers [21], nucleic acid dendrimers [21] and poly(lysine) dendrimers [23, 24] (see Figures 10.2, 10.4 and 10.6). However, in each case appropriate water solubilizing terminal groups are required (i.e. -NH2, -OH or C02H groups) for suitable electrophoretic analysis. 

Allen, R. C. Gel Electrophoresis of Proteins and Nucleic Acids Selected Techniques, W. de Gruyter, New York, 1994. 

To understand how these modem methods work, it is necessary first to review some general laboratory techniques ubiquitous in genetic engineering. Among the most important are gel electrophoresis of nucleic acids, nucleic acid hybridization assays, and the polymerase chain reaction. 

Gel Electrophoresis. This is becoming a more commonly used procedure for purifying proteins, nucleic acids, nucleoproteins, polysaccharides and carbohydrates. The gels can be electroblotted onto membranes and the modem procedures of identifying, sequencing (proteins and nucleic acids) and amplifying (nucleic acids) on sub-micro scales have made this technique of separation a very important one. (See D.Patel Gel Electrophoresis, J.Wiley-Lis, Inc., 1994). 

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