DNA analysis, by capillary

Muller, O., Minarik, M., and Foret, F., Ultrafast DNA analysis by capillary Electrophoresis/laser-induced fluorescence detection, Electrophoresis, 19,1436, 1998. 

Righetti PG, Gelfi C, D Acunto MR. Recent progress in DNA analysis by capillary electorphoresis. Electrophoresis 2002 23 1361-1374. 

Fasco MJ, Treanor CP, Spivack S, Figge HL, Kaminsky LS (1995) Quantitative RNA-polymerase chain reaction—DNA analysis by capillary electrophoresis and laser induced fluorescence. Anal Biochem 224 140-147. 

Gao, Q., Shi, Y., and Liu, S., Multiple-channel microchips for high-throughput DNA analysis by capillary electrophoresis, Fresenius J. Anal. Chem., 371, 137, 2001. 

Apruzesse, W.A. and Vouros, P, Analysis of DNA adducts by capillary methods coupled to mass spectrometry a perspective, /. Chromatogr. A, 794, 97, 1998. 

I. Hoes, E.L. Esmans, Analysis of melphalan adducts of 2 -deoxynucleotides in calf thymus DNA hydrolysates by capillary LC-ESI-MS-MS, J. Chromatogr. B, 736 (1999)43. 

The limitations of microtechnology are invariably related to the concentration of the analyte or type of cell under study. If the device is only capable, of receiving microliter quantities, then the final signal strength being measured will depend on the inclusion of an amplification process (e.g., PCR) or the ability to detect extremely low levels of analyte concentration or type of cell identification. For example, only O.SpL of whole blood are necessary to ahow for isolation of 500 WBCs, more than sufficient to provide genomic DNA for mutation detection by PCR. Similarly, submicroliter quantities of protein or DNA solutions provide adequate material for analysis by capillary electrophoresis (see Chapter 5). However, if the aim is to identify and isolate an infected WBC m whole blood that is present at an incidence of only 1 in 10 million, then quantities in excess of 10 mL of whole blood may be required just to encounter 5 cells. This does not provide the ideal situation for a microdevice. Another key limitation, namely the impact of surface chemistry, has been addressed in a previous section. 

Giovannoli, C., Anfossi, L., Tozzi C., Giraudi, G, and Vanni, A. DNA separation by capillary electrophoresis with hydrophilic substituted celluloses as coating and sieving polymers. Apphcation to the analysis of genetically modified meals, J. Sep. ScL, 27, 1551, 2004. 

Wang, Y, et al.. Quasi-interpenetrating network formed by polyacrylamide and poly(lV,lV-dimethylacrylamide) used in high-performance DNA sequencing analysis by capillary electrophoresis, Electrophoresis, 26, 126, 2005. 

Salas-Solano, O., Ruiz-Martinez, M.C., Carrilho, E., Kotler, L., and Karger, B.L., A sample purification method for ragged and high-performance DNA sequencing by capillary electrophoresis using replaceable polymer solutions. B. Quantitative determination of the role of sample matrix components on sequencing analysis, AnoZ. Chem., 70, 1528, 1998. 

Atha, D., Kasprzak, W., O Connell, C., etal.. Prediction of DNA single-strand conformation polymorphism Analysis by capillary electrophoresis and computerized DNA modeling. Nucleic Acids Res, 29, 4643, 2001. 

Fasco, M. J. Analysis of amplified DNA molecules by capillary electrophoresis and laser induced fluorescence. Methods Mol. Med. 1999,26,131-146. 

Fig. 1. Southern blot analysis of DNA showing (a) step 1, an agarose gel containing separated restriction fragments of DNA, denoted by (—), which is immersed in NaOH to denature the double-stranded stmcture of DNA, and then transferred by capillary flow to a nitrocellulose filter. In step 2, the bound DNA is allowed to hybridize to a labeled nucleic acid probe, and the unbound probe is washed off In step 3, the filter is placed into contact with x-ray film resulting in (b) bands of exposure on the film which are detected after development and correspond to regions where the restriction fragment is...

Arakawa, H., Nakashiro, S., Maeda, M., and Tsuji, A., Analysis of single-strand DNA conformation polymorphism by capillary electrophoresis, /. Chromatogr. A, 722, 359, 1996. 

Method validation depends on the type and purpose of analysis. For example, the recommended validation procedure for PCR, followed by capillary gel electrophoresis of recombinant DNA, may consist of the following steps ... 

SNP arrays result useful to detect DNA rearrangements by an indirect approach. In this case, the result will be measured as a loss of heterozygosity, a statistical value which measures DNA deletions due to statistical deviations from the expected rate in the different alleles. Loss of heterozygosity can also be measured at a shorter scale by capillary electrophoresis, in this case restricted to individual gene analysis (92). 

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