Methylation specific PCR is reproducible within a wide dynamic range.
<p><b><b>A</b></b> Both detection of a single copy gene and rDNA are linear between 0.3 ng and 20 ng per reaction. The single copy gene ACTB is insensitive to methylation status and detects the same copies/µL with and without M.SssI treatment. Methylated rDNA has lower...
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2025
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| Summary: | <p><b><b>A</b></b> Both detection of a single copy gene and rDNA are linear between 0.3 ng and 20 ng per reaction. The single copy gene ACTB is insensitive to methylation status and detects the same copies/µL with and without M.SssI treatment. Methylated rDNA has lower copies/µL than total rDNA as expected. DNA input is for a 12 µL dPCR reaction with the number of copies per µL indicating the absolute quantification from dPCR. <b><b>B</b></b> When normalized to ACTB, rDNA tCN and mCN are stable across our detection range with mCN consistently less than tCN. rDNA CN reflects normalization to ACTB independently for native (mCN) and M.SssI treated (tCN) aliquots. When compared to theoretical genome copies/ µL we report < 15% error consistently with an average percent error of 7.5%.</p> |
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