Journal: Nucleic Acids Research

Article Title: Methylation enrichment pyrosequencing: combining the specificity of MSP with validation by pyrosequencing

doi: 10.1093/nar/gkl424

Figure Lengend Snippet: Diagrammatic representation of primer alignment on sodium bisulphite-treated p16 promoter sequence. Cytosines in squares indicate the nucleotides interrogated by pyrosequencing in PMA and MEP assays (i.e. CpG dinucleotides). The T nucleotide in a square indicates the bisulphite control of these assays (site of non-CpG cytosine). MEP, methylation enrichment PCR primers; PMA, pyrosequencing methylation assay primers; PSEQ, pyrosequencing primer; MSP, methylation-specific PCR primers (primers specific for unmethylated target DNA contain T in squares; primers specific for methylated DNA target contain C in squares).

Article Snippet: We describe a novel combination of techniques conferring the sensitivity and specificity of MSP while also benefiting from subsequent validation using pyrosequencing: methylation enrichment pyrosequencing (MEP).

Techniques: Sequencing, Control, Methylation

Journal: Nucleic Acids Research

Article Title: Methylation enrichment pyrosequencing: combining the specificity of MSP with validation by pyrosequencing

doi: 10.1093/nar/gkl424

Figure Lengend Snippet: Tumour characteristics and p16/cyclin A1 methylation results

Article Snippet: We describe a novel combination of techniques conferring the sensitivity and specificity of MSP while also benefiting from subsequent validation using pyrosequencing: methylation enrichment pyrosequencing (MEP).

Techniques: Methylation

Journal: Nucleic Acids Research

Article Title: Methylation enrichment pyrosequencing: combining the specificity of MSP with validation by pyrosequencing

doi: 10.1093/nar/gkl424

Figure Lengend Snippet: Agarose gels of PCR products following ( a ) p16 MSP, ( b ) p16 MEP, and ( c ) cyclin A1 MEP. U, PCR using MSP primers specific for unmethylated DNA; M, PCR using methylation-specific MSP primers; A and B, two independent MEP assays. Rows 1–4 (top to bottom) correlate with rows 1–4 in dilution matrix ; i.e. row 1, no dilution of starting M:U concentrations; row 2, 1/10 dilutions, etc. Lanes 1–8 correlate with columns 1–8 in dilution matrix ; i.e. lane 1, M:U ratio of 1:1; lane 2, M:U ratio of 1:5, etc.

Article Snippet: We describe a novel combination of techniques conferring the sensitivity and specificity of MSP while also benefiting from subsequent validation using pyrosequencing: methylation enrichment pyrosequencing (MEP).

Techniques: Methylation

Journal: Nucleic Acids Research

Article Title: Methylation enrichment pyrosequencing: combining the specificity of MSP with validation by pyrosequencing

doi: 10.1093/nar/gkl424

Figure Lengend Snippet: Pyrogram for a typical p16 MEP showing 100% methylation. Shaded bars encompassing T/C pairs, seven interrogated CpG (T, conversion of unmethylated C to U; C, non-conversion of methylated C); box, control, non-CpG cytosine (0% cytosine incorporated). a: Sequence context ATYG. Equal height of T and C peaks at 5 units indicates 100% methylation of C. b: Sequence context GTTYGG. Height of T peak (10 units) is twice the height of C peak (5 units) indicating 100% methylation.

Article Snippet: We describe a novel combination of techniques conferring the sensitivity and specificity of MSP while also benefiting from subsequent validation using pyrosequencing: methylation enrichment pyrosequencing (MEP).

Techniques: Methylation, Control, Sequencing