THE FEASIBILITY OF IN SITU BISULFITE MODIFICATION FOR GENOMIC DNA METHYLATION DETECTION
With the increasing interest in DNA methylation, array-based methods have been developed recently. However, in these methods, the complex sample treatment processes are necessary, such as bisulfite modification in liquid state of every sample. Here, we developed a micro array-based technology to interrogate DNA methylatton status, in which the DNA samples were firstly dotted on solid support, treated with bisulfite directly and hybridized. In this assay, plasmid pUC19 DNA was immobilized on nylon membranes and Acrylamide Gel chips respectively, and soaked in bisulfite reaction mixtures to convert unmethylated Cs to Us, and methylated Cs unchanged. The probes for detection of the methylation of CpG sites were hybridized with the converted DNA. Here two immobilized methods were used to indicate that to immobilized DNA on solid support for in situ bisulfite modification to detect DNA methylation is a feasible method. From the result of these two methods, detecting DNA methylation without PCR of in situ bisulfite modification is suitable for the small genomic DNA. The experiments show that this method can simplify the experiment processes and increase the efficiency of the bisulfite treatment.
DNA methylation sodium bisulfite treatment dot blot acrylamide gel
Yan Wang Yuan Wan Wenli Zheng Junfeng Luo Dingdong Zhang Zuhong Lu
The State Key Laboratory of Bioelectronics, Department of Biological Science and Medical Engineering,Southeast University, Nanjing 210096, China
国际会议
The 4th International Forum on Post-genome Technologies(4IFPT)(第四届国际后基因组生命科学技术学术论坛)
杭州
英文
173-177
2006-09-25（万方平台首次上网日期，不代表论文的发表时间）