Establishment of restriction landmark genome scanning for detection of genomic DNA methylation of tissues
- VernacularTitle:限制性酶切扫描技术测定DNA甲基化组的方法建立
- Author:
Yun QIN
;
Da-Jun DENG
;
Li YU
;
- Publication Type:Journal Article
- Keywords:
Neoplasms;
DNA methylation;
DNA restriction landmark genome scanning
- From:
Chinese Journal of Laboratory Medicine
2003;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
50 000 bp)was extracted from frozen gastric cancer tissue and their corresponding normal samples and used for RLGS assay.The genome DNA was digested by methylation-sensitive restriction enzyme Not Ⅰ, and labeled by radioisotope ~(32)p,then separated by two-dimensional electrophoresis and autoradiography. Experimental conditions for each step were optimized step by step.DNA fragment sequences for the dots on scanning profile were identified according to the human RLGS database.Results RLGS assay was set up and used to get the RLGS profiles of the representative tested samples successfully.These profiles can display more than 1 200 available dots averagely,the profiles of high quality DNA sample can display more than 1 800 dots which is the average level at an excellent RLGS lab,discrepant dots which had weaken or enhanced signals and their sequence information were obtained.The result can be reproduced.Conclusion The RLGS assay is established,stabilized for detection of DNA methylation of tissue samples.
