甲基化分析

I am planning to perform sodium bisulphite sequencing. Currently I am trying to design primers of the promoter region of my target genes. However some of these promoters have not been well-characteriz ...

S1 Nuclease Protection Assay End-label oligo (20-25 mer) 4 µl 5X T4 Kinase Buffer 5 µl ATP (7000 Ci/mmol) 10 µl water + oligo (200 ng) 1 µl T4 Kinase 1. 37 deg C for 1 hour. ...

DNA methylation is an epigenetic event that affects cell function by altering gene expression and refers to the covalent addition of a methyl group catalyzed by DNA methyltransferase (DNMT) to the 5-c ...

Source: Protocol Online Abstract: Modifying DNA using sodium bisulfite to convert unmethylated cytosines to uracils and subsequently detect methylated cytosines using methylation specific PCR (MSP) te ...

Pyrosequencing 在肿瘤基因甲基化研究的应用 甲基化研究对于基因的调控和肿瘤的发生有非常密切的关系。在一般正常的细胞中，基因调控区CpG岛处于非甲基化的状态。而当细胞发生癌变后，这些CG区域往往呈现甲基化状态。英国医学刊物《Lancet》报道奥地利因斯布鲁克医学院的专家们早就可以通过检测大便中DNA的甲基化变化，把健康人的大便与结肠癌患者的大便区分开。大便标本中SFRP2 甲基化的程 ...

The restriction/modification system in bacteria is a small-scale immune system for protection from infection by foreign DNA. W. Arber and S. Linn (1969) Plating efficiencies of bacteriophage lambda (l ...

Adapted from Frommer et.al.* 1.Dilute DNA (up to 2 mg) into 50 ml with distilled H2O. 2.Add 5.5 ml of 2M NaOH. 3.Incubate at 37℃ for 10 minutes (to create single stranded DNA ). 4.Add 30 ml of 10 ...

Footprinting Footprinting is a method for determining the exact DNA sequence to which a particular DNA-binding protein binds. Examples: hormone-receptor complexes that bind to their hormone response elements transcription factors that bind eukaryotic operators, enhancers, and silencers the lac repressor that shuts down the lac operon in E. coli [Discussion of the lac operon] How, for example, does one determine the DNA sequence to which the lac repressor binds? The procedure: Clon