Denaturing high performance

A series of papers with first systematic studies of SNPs in selected genes have been published recently (Cargill et al., 1999 Halushka et al., 1999, Hacia et al., 1999). In all studies high-density variant detector arrays (Chee et al., 1996 Lipshutz et al., 1999) were used to detect SNPs in PCR products. In the study denaturing high-performance liquid chroma-... 

A.R. Bahrami, M.J. Dickman, M.M. Matin, J.R. Ashby, P.E. Brown, M.J. Conroy, G.J.S. Fowler, J.P. Rose, Q.I. Sheikh, A.T. Yeung and D.P. Hornby, Use of fluorescent DNA-intercalating dyes in the analysis of DNA via ion-pair reversed-phase denaturing high-performance liquid chromatography. Anal. Biochem., 309 (2002) 248-252. 

Ul. Underhill, P. A., Jin, L., et al. Detection of numerous Y chromosome biallelic polymorphisms by denaturing high-performance liquid chromatography. Genome Res. 7(10), 996-1005 (1997). 

Fischer J, Sehwab M, Eichelbaum M et al. Mutational analysis of the human dihydropyrimidine dehydrogenase gene by denaturing high-performance liquid chromatography. Genet Teit 2003 7 97-105. 

Liu W, Smith DI, Rechtzigel KJ, Thibodeau SN, James CD (1998) Denaturing high performance liquid chromatography (DHPLC) used in the detection of germline and somatic mutations. Nucleic Acids Res 26 1396-1400... 

Giordano M, Oefner PJ, Underhill PA, Cavalli-Sforza LL, Tosi R, Richiardi PM. Identification by denaturing high-performance liquid chromatography of numerous polymorphisms in a candidate region for multiple sclerosis susceptibility. Genomics 1999 56 247-253. 

Iwamoto K, Kato T. Effects of cocaine and reserpine administration on RNA editing of rat 5-HT2C receptor estimated by primer extension combined with denaturing high-performance liquid chromatography. Pharmacogenomics J 2002 2 335-340. 

FIGURE 16.1 Steps of denaturing high performance liquid chromatography (DHPLC). 

Lam, C.W. Analysis of polymerase chain reaction products by denaturing high-performance liquid chromatography. Meth. Mol. Biol. 2006, 336, 73-82. 

Gross, E. et al. Allelic loss analysis by denaturing high-performance liquid chromatography and electrospray ionization mass spectrometry. Human Mutat. 2007, 28, 303-311. 

Figure 3. Denaturated high performance liquid chromatography of DNA fragments including a SNP. The fragment of a sample is always mixed with a reference isolate without SNP.

Xiao M, Oefner PJ. Denaturing high performance chromatography a review. Hum Mutat 2001 17 439-74. 

Franciosa, G., Pourshaban, M., De Luca, A., Buccino, A., Dallapiccola, B. and Aureli, P Identification and Type A, B, E, and F botulinum neurotoxin genes and of botulinum neurotoxigenic Clostridia by denaturing high-performance liquid chromatography. Appl. Environ. Microbiol., 70, 4170-4176 (2004). 

SSCP, single-strand conformation polymorphism PNA-LNA, peptide nucleic acid-locked nucleic acid MALDI-TOF MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry ARMS, amplified refractory mutation system dHPLC, denaturing high performance liquid chromatography. 

One specialty in dehybridization detection is that of dHPLC (denaturing high-performance liquid chromatography). Hybridization products differ in melting temperature and mobility on a specific support if a mismatch is introduced by a SNP. In the case of a SNP, a heteroduplex is formed in the analyzed sequence, whereas in the case of identical sequences (no SNP present), a perfect homoduplex is formed. These events can be discriminated by dHPLC at a low to medium throughput for SNP discovery or SNP detection. 

D-HPLC, see denaturating high-performance liquid chromatography DHFR (Dehydrofolate reductase) 725, 1079... 

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