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Image Search Results
Journal: Nucleic Acids Research
Article Title: Intra-tumor heterogeneity of MLH1 promoter methylation revealed by deep single molecule bisulfite sequencing
doi: 10.1093/nar/gkp457
Figure Lengend Snippet: Comparison of sequencing results from conventional cloning and Sanger sequencing and 454 Life Sciences FLX sequencing of MLH1 promoter PCR products from bisulfite treated DNA of an endometrial cancer and normal blood from Patient #1684. ( A ) Schematic of the MLH1 promoter is presented with arrows indicating the location of PCR primers and vertical lines representing the position of CG dinucleotides. Below the schematic are the results from cloning and bisulfite sequencing 45 molecules from the distal (left) and proximal (right) promoter in each sample (tumor and matched normal blood). Each column represents a CG dinucleotide in the sequence, and corresponds to each vertical line in the promoter schematic. Each row represents a single molecule. The color of the boxes represents the methylation state of each cytosine. Red, methylated; Black, unmethylated. ( B ) The results from the FLX single molecule sequencing of the same samples. The distance of each CpG from the transcription start site (UCSC Human Genome March 2006) is listed from distal to proximal. The Distal Amplicon: −671, −662, −654, −648, −634, −632, −630, −626, −623, −619, −609, −605, −596, −584, −576, −569, −566, −564, −560, −558, −548, −540, −537, −512, −505, −483, −470, −465, −449, −446, −421, −405, −380, −368. The Proximal Amplicon: −341, −325, −318, −286, −280, −249, −240, −226, −209, −202, −192, −190, −184, −165, −154, −139, −70, −47, −23, −15. As an additional point of reference, the translation start site (ATG) is currently annotated at +60 downstream of the transcription start site (position 0).
Article Snippet: Equimolar quantities of each PCR product were then pooled into a single tube, purified on a Qiaquick column (Qiagen Inc, Valencia, CA) and submitted to Cogenics Inc. (
Techniques: Sequencing, Clone Assay, Methylation Sequencing, Methylation, Amplification