DNA Mutation Detection Using Denaturing High‐Performance Liquid Chromatography (DHPLC)

Yu, Bing; Sawyer, Nicole; Chiu, Christine L; Oefner, Peter J.; Underhill, Peter A.

doi:10.1002/0471142905.hg0710s19

Cited by 143 publications

(165 citation statements)

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“…13 -16 The presence of the Y Alu polymorphic insertion was tested as described elsewhere. 17 Genotyping was done by using the denaturing high performance liquid chromatography method proposed by Oefner and Underhill, 18 with a phylogenetic hierarchical approach. The V12, V13, V22 polymorphisms, defining Hgs E3b1a1, E3b1a2 and E3b1a3, have been analysed as described in Cruciani et al 16 Data are referred to terminal mutation and according to the International Society of Genetic Genealogy nomenclature.…”

Section: Methodsmentioning

confidence: 99%

Differential Greek and northern African migrations to Sicily are supported by genetic evidence from the Y chromosome

et al. 2008

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Section: Methodsmentioning

confidence: 99%

Differential Greek and northern African migrations to Sicily are supported by genetic evidence from the Y chromosome

et al. 2008

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“…Seperation of heteroduplex molecules from homoduplex molecules is performed by ion-pair, reverse phase liquid chromatography. 19,20 DHPLC analysis has been performed after mixing tumor samples and wild-type DNA at a ratio of 2:1. The PCR reaction was performed in 25 ll of a reaction mixture consisting of 10 mM Tris-HCl (pH 5 8.3), 50 mM KCl, 1.0, 1.5 or 2.0 mM MgCl 2 , 0.01% gelatin, and 200 mM dNTP 0.4 mM of previously published primers.…”

Section: Analysis Of P53 Mutationsmentioning

confidence: 99%

The prognostic impact of O⁶‐Methylguanine‐DNA Methyltransferase (MGMT) promotor hypermethylation in esophageal adenocarcinoma

Baumann

Keller

Pühringer

et al. 2006

Intl Journal of Cancer

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Promotor hypermethylation is a common event in human cancer. O 6 -Methylguanine-DNA Methyltransferase (MGMT) is a gene involved in DNA repair, which is methylated in a variety of cancer types. In colorectal cancer and lung cancer, hypermethylation of MGMT has been correlated with p53 mutation. In the present study, 132 samples of esophageal adenocarcinoma and 58 samples of normal esophageal tissue were investigated for MGMT hypermethylation status by methylation-specific real-time PCR and results were correlated to clinicopathological parameters, patient's survival, p53 mutation and expression of p53 protein and MGMT protein. In the carcinomas, hypermethylation of MGMT was found in 63.6% of cases and loss of MGMT protein expression in 48.5% of cases. Furthermore, MGMT hypermethylation was found in 5.7% of normal esophageal smooth muscle tissue, in 20.0% of esophageal squamous epithelium and in 61.5% of nonneoplastic Barrett's mucosa. In the carcinomas, hypermethylation of the MGMT gene was correlated with loss MGMT protein expression (p < 0.0001) and with high tumor differentiation (p 5 0.0079). In contrast, no correlation between MGMT hypermethylation, Lauren's classification, WHO classification, tumor size, gender, age, pT category and pN category, and p53 status was found. Neither MGMT hypermethylation nor loss of MGMT protein expression was correlated with patient's survival. In conclusion, MGMT hypermethylation in esophageal adenocarcinoma is a frequent event that is associated with loss of MGMT protein expression but not with patient's outcome. ' 2006 Wiley-Liss, Inc.Key words: esophageal adenocarcinoma; MGMT; hypermethylation; prognosis; p53During the last few years, cancer researchers have focused on methylation of cytosine residues within a CpG island. Methylation of cytosine per se is not only found in cancer cells but also in all normal cells in which 3-4% of cytosines are methylated. However, methylation of CpG islands in normal cells occurs only in a limited number of conditions, e.g. in the case of gene imprinting. 1 Since CpG islands are mostly located in promotor regions, i.e. the 5 0 untranslated region or the exon 1 of a gene, CpG island methylation often has an influence on gene expression. 2 Promotor methylation of tumor supressor genes or DNA repair genes has been found within various types of cancer. Additionally it has been shown that promotor methylation of certain genes may be tumor specific and tissue specific. [3][4][5][6] One gene whose promotor has been found methylated in 20-30% of human tumor cell lines is that of the enzyme O 6 -methylguanine-DNA methyltransferase (MGMT; E.C. 2.1.1.63). 7 MGMT, a DNA repair enzyme, removes methyl-or alkyl-groups from guanine after chemical modification and therefore protects cells from G to A mutations. 8 For that reason, hypermethylation of the MGMT promotor can promote the accumulation of mutations within the cell. Previous investigations indicated that MGMT hypermethylation can facilitate G:C to A:T mutations in the tumor suppressor gene p53 i...

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“…To identify variation in the TRAIL gene, we screened all five exons, including the 5Ј and 3Ј untranslated regions, in PCR-amplified DNA from 10 Caucasian and 10 African American individuals by denaturing high performance liquid chromatography (DHPLC). 9 All PCR fragments that showed variant DHPLC patterns were directly sequenced (Figure 1). …”

mentioning

confidence: 99%

Three polymorphisms in the 3′ UTR of the TRAIL (TNF-related apoptosis-inducing ligand) gene

et al. 2001

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DNA Mutation Detection Using Denaturing High‐Performance Liquid Chromatography (DHPLC)

Cited by 143 publications

References 21 publications

Differential Greek and northern African migrations to Sicily are supported by genetic evidence from the Y chromosome

Differential Greek and northern African migrations to Sicily are supported by genetic evidence from the Y chromosome

The prognostic impact of O⁶‐Methylguanine‐DNA Methyltransferase (MGMT) promotor hypermethylation in esophageal adenocarcinoma

Three polymorphisms in the 3′ UTR of the TRAIL (TNF-related apoptosis-inducing ligand) gene

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DNA Mutation Detection Using Denaturing High‐Performance Liquid Chromatography (DHPLC)

Cited by 143 publications

References 21 publications

Differential Greek and northern African migrations to Sicily are supported by genetic evidence from the Y chromosome

Differential Greek and northern African migrations to Sicily are supported by genetic evidence from the Y chromosome

The prognostic impact of O6‐Methylguanine‐DNA Methyltransferase (MGMT) promotor hypermethylation in esophageal adenocarcinoma

Three polymorphisms in the 3′ UTR of the TRAIL (TNF-related apoptosis-inducing ligand) gene

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The prognostic impact of O⁶‐Methylguanine‐DNA Methyltransferase (MGMT) promotor hypermethylation in esophageal adenocarcinoma