PCR Methylation specific PCR - Mammalian DNA

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.

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1 year ago

1 year ago by Daniel McKenzie United Kingdom

How “strong” is the PCR product of methylation specific PCR?

Does anyone know how “strong” the PCR product of methylation specific PCR is? I kept my PCR products at 4C for about 3 weeks and then at room temperature for another week. Will I be able to use them for sequencing?

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Protocol tips
For best results, the CT Conversion Reagent should be prepared freshly and used immediately following preparation.
Protocol tips
For best results, the CT Conversion Reagent should be prepared freshly and used immediately following preparation.
Upstream tips
Aliquote in small quantity to avoid freeze-thaw cycles and Storage at -70°C for long-term use (2-3 yrs)
Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.
Protocol tips
Follow manufacturer’s instructions
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