[The multiplex analysis of epigenetic markers and genetic markers by post-digestion mutagenically separated PCR].
OBJECTIVE
To establish a novel method for the multiplex analysis of the methylation and single nucleotide polymorphism (SNP).


METHODS
The imprinted SNP rs220028 was chosen as a model. Genomic DNA, after being digested with methylation sensitive restriction enzyme, were typed by mutagenically separated PCR (MS-PCR). The polymorphism of restriction site was excluded by PCR-RFLP.


RESULTS
By post-digestion MS-PCR, the methylated allele was detected selectively, the maternal origin of which was confirmed by pedigree analysis; A=0.5085, G=0.4915,PIC=0.3749.


CONCLUSIONS
The multiplex analysis of methylation markers and SNP can be achieved by post-digestion MS-PCR. The imprinted SNP locus rs220028 is a potentially useful marker in screening Prader-Willi/Angelman syndrome.
Zhao
Gui-sen
GS
Department of Forensic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030 PR China.
Huang
Dai-xin
DX
Feng
Wen-fang
WF
Yang
Qing-en
QE
D019175:DNA Methylation; D004262:DNA Restriction Enzymes / Q000378:metabolism; D005819:Genetic Markers / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide
D005819:Genetic Markers; D004262:DNA Restriction Enzymes