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Bisulfite Conversion and PCR Amplification of DNA Sequences for Methylation Analysis

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  1. H19 DMR 5’ 3’ GTAGATTTGGTTATAGTTAAATGGATA----ATTTTATAGTTATAAAAGTAGAGGTTGG CATCTAAACCAATATCAATTTACCTAT----TAAAATATCAATATTTTCATCTCCAACC 3’ 5’ PCR 5’ 3’ GTAGATTTGGTTATAGTTAAATGGATA----ATTTTATAGTTATAAAAGTAGAGGTTGG Converted top strand Bisulfite 5’ 3’ GCAGATTTGGCTATAGCTAAATGGACA----ACCCCATAGCCATAAAAGCAGAGGCTGG CGTCTAAACCGATATCGATTTACCTGT----TGGGGTATCGGTATTTTCGTCTCCGACC 3’ 5’ (Beware of CpG dinucleotides in primer sequence) Bisulfite TGTTTAAATTGATATTGATTTATTTGT----TGGGGTATTGGTATTTTTGTTTTTGATT 3’ 5’ Converted bottom strand PCR 5’ 3’ ACAAATTTAACTATAACTAAATAAACA----ACCCCATAACCATAAAAACAAAAACTAA TGTTTAAATTGATATTGATTTATTTGT----TGGGGTATTGGTATTTTTGTTTTTGATT 3’ 5’

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