HIGH-THROUGHPUT DETECTION OF p16Ink4a PROMOTER METHYLATION USING ACRYLAMTOE-MODIFIED NUCLEIC ACID MICROARRAY
In recent years, DNA microarray methods have been developed for detecting DNA methylation. The current chemoimmobilization methods merely attach a small quantify of oligonucleotide probes or PCR products on the glass slide that lower the sensitivity of detection. Further, researchers have to purify a large amount of PCR products to improve the efficiency and stability of DNA attachment. We utilized three-dimensional functionalized hydrophilic polyacrylamtde gel to heighten the quantity of immobilized DNA. In addition, electrophoresis instead of washing was adopted in order to effectively remove the non-specifically bound probes in the post-hybridization process. In our experiments, we showed that p16 was methylated in the positive control, while no methylation was found in the negative control. The results were further validated by methylation-specific PCR and bisulfite DNA sequencing. The coupling of acrylamide-modified nucleic acid microarray with linker-PCR could be used as an inexpensive, convenient and sensitive tool for detection DNA methylation of multiple samples in the future.
DNA microarray methylation acrylamide gel linker-PCR,p16
Yuan Wan Yan Wang Junfeng Luo Zuhong Lu
State Key Laboratory of Bioelectronics, 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
118-121
2006-09-25（万方平台首次上网日期，不代表论文的发表时间）