Details

Autor(en) / Beteiligte

• Couvert, P
• Poirier, K
• Carrié, A
• Chalas, C
• Jouannet, P
• Beldjord, C
• Bienvenu, T
• Chelly, J
• Kerjean, A

Titel

DHPLC-Based Method for DNA Methylation Analysis of Differential Methylated Regions from Imprinted Genes

Ist Teil von

• BioTechniques, 2003-02, Vol.34 (2), p.356-362

Ort / Verlag

Natick, MA: Future Science Ltd

Erscheinungsjahr

2003

Quelle

MEDLINE

Beschreibungen/Notizen

• The bisulfite genomic sequencing method is one of the most widely used techniques for methylation analysis in heterogeneous unbiased PCR, amplifying for both methylated and unmethylated alleles simultaneously. However, it requires labor-intensive and time-consuming cloning and sequencing steps. In the current study, we used a denaturing high-performance liquid chromatography (DHPLC) procedure in a complementary way with the bisulfite genomic sequencing to analyze the methylation of differentially methylated regions (DMRs) of imprinted genes. We showed reliable and reproducible results in distinguishing overall methylation profiles of DMRs regions of human , , , , , , , and mouse , , imprinted genes. These DHPLC profiles were in accordance with bisulfite genomic sequencing data and may serve as a type of "fingerprint," revealing the overall methylation status of DMRs associated with sample heterogeneity. We conclude that DHPLC analysis could be used to increase the throughput efficiency of methylation pattern analysis of imprinted genes after the bisulfite conversion of genomic DNA and unbiased PCR amplification.