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[Analysis of demethylation-related HPV16 reactivation by DHPLC-primer extension assay].
Zhonghua Yu Fang Yi Xue Za Zhi
June 2007
Peking University School of Oncology and Beijing Institute for Cancer Research, Beijing 100036, China.
Objective: To establish a sensitive assay to detect methylation status of the critical 7862nt CpG site related to transcription of HPV16 E6 and E7 genes.
Methods: Genomic DNA of two HPV16-infected cell line SiHa and CaSki was extracted and modified by sodium bisulfite to convert the unmethylated Cs to Us (Ts in PCR products). The target sequence of HPV16 including the 7862nt CpG site was pre-amplified by PCR.
Multiple molecular analyses from minimal cell quantities by sequential isolation and preamplification of DNA and RNA.
Diagn Mol Pathol
September 2007
Department of Pathology and Molecular Diagnostics, University of Regensburg, Germany.
Molecular analyses of single or very few cells are often handicapped by low amounts of DNA or RNA from starting material. Specimens like biopsies, embryonic stem cells, or laser-microdissected tissues often do not provide nucleic acid quantities required for robust amplification and verification by diagnostic polymerase chain reaction (PCR)-based analyses. A simultaneous isolation procedure for both RNA and DNA from a single sample would greatly facilitate demanding molecular studies.
View Article and Find Full Text PDFGenotyping of three single nucleotide polymorphisms following whole genome preamplification of DNA collected from buccal cells.
Behav Genet
September 2004
Institute for Behavioral Genetics, University of Colorado, Boulder, CO 80309-0447, USA.
Collection of genomic DNA from buccal cells is a simple and convenient procedure for genotyping individuals. One disadvantage is that the amount of genomic DNA may be inadequate for genotyping projects that require a large number of determinations per sample. Primer Extension Preamplification (PEP) methods that can amplify the entire genome 100-fold or more offer a potential solution to this problem.
View Article and Find Full Text PDFAlterations in p53 predict response to preoperative high dose chemotherapy in patients with gastric cancer.
Mol Pathol
October 2003
Department of Pathology, University of Regensburg, 93042 Regensburg, Germany.
Aims: To evaluate the usefulness of molecular markers in predicting histopathological and clinical response to preoperative high dose chemotherapy (HDCT) and survival of patients with advanced gastric cancer.
Methods: In a phase II trial, 25 patients with metastatic gastric cancer received preoperative tandem HDCT consisting of etoposide, cisplatin, and mitomycin, followed by autologous bone marrow transplantation to achieve surgical resectability. Samples before and after treatment, from normal and tumour tissue, were characterised histopathologically, and both p53 and BAX expression was analysed by immunohistochemistry.
Primer-extension preamplified DNA is a reliable template for genotyping.
Clin Chem
September 2002
Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, MI 48201-1928, USA.
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