【摘 要】
：
Objective To develop a quantitative method for methylation analysis of the p16 gene based on mismatch hybridization and chemiluminescence. Methods Genomic DNA w
【机 构】
：
Institute of Environmental Medicine
论文部分内容阅读
Objective To develop a quantitative method for methylation analysis of the p16 gene based on mismatch hybridization and chemiluminescence. Methods Genomic DNA was modified by sodium bisulfite to convert all unmethylated but not methylated cytosines to uracil, and subsequently a pair of primer having no CpG sites was designed for amplification target DNA containing methylated or unmethylated CpG sites. The PCR product spanning CpG sites were hybridized with two oligonucleotide probes which perfectly matched the methylated and unmethylated CpG sequences respectively, and the hybrids were detected by chemiluminescent method. The percentage of methylated target sequences could be estimated by calculating the ratio of signals obtained with two probes. Results The percentage of methylation of artificial mixtures DNA showed a linear relation. There was a negative correlation between the methyaltion index with p16 transcriptional mRNA of p16 gene in tumor cell lines. Conclusion Compared with existing methods, this assay is nonisotopic, rapid, simple, and can be widely applied to the study of DNA methylation.
其他文献
In-flight particle velocity and flux distribution were measured using CCD thermal spray monitor system during supersonic plasma spray processing with nano-struc
The temperature dependence of surface enhanced Raman scattering of the C70 molecule is reported.The Raman scattering of C70 molecules adsorbed on the surface of
利用改进的量子分子动力学研究了16O+16O库仑位垒附近的熔合反应,用连接函数的方法对体系的表面能做了改进.利用这种方法避免了非物理的核子发射,熔合截面有了明显的增加,这
A new centrifuge based method for determining the response of continuous buried pipe to PGD is presented. The physical characteristics of the RPI’s 100 g-ton g
In this study, we determined pH, phosphorus content and the number of P-dis- solving/decomposing bacteria in relation to the depth in the sediment of Guanting r
As a type of chemical adsorption working pair, the physical adsorption occurs first for CaCl2-NH3 because the effective reaction distance for van der Waals forc
Poly(tetrafluoroethylene-co-hexafluoropropylene) (FEP) films were immersed in aqueous solution of acrylic acid (AA) and sodium styrene sulfonate (SSS), then irr
Using vapor transport equilibration (VTE) technique we succeeded in the fabrication of single-phase γ-LiAlO2 layer on (0001) sapphire substrate. X-ray diffract
By utilizing observational data from a 325 m tower of the Institute of Atmospheric Physics (IAP), Chinese Academy of Sciences (CAS) on March 19-29, 2001 and Aug
Localized nature of damage in structures requires local measurements for structural health monitoring. The local measurement means to measure the local, usually