I did arms pcr using sybr green real time pcr.. I got an earlier ct for one of the primers but higher end point for the other primer.. what can I do?

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Soner Öner-Sieben

Hi Ahmed,

sequencing your amplicons would be a good idea.

Best regards

Ahmed Fakhr

Hi Soner,

Thank you for your answer.

But I think in ARMS PCR I used a 2 forward primers with a single base difference against a universal reverse primer in 2 separate reactions. If I make sequencing using the common reverse primer I think these last bases would not be good to analyse. In addition I think I will get the base in the used primer in each amplicon.

I would clone the amplicon into a vector like pGEM-T and than ship it for sequencing. With that, sequencing can be done we standard primers like SP6 and T7 flanking your sequence in the vector.

The actual problem is that I am doubting the results of ARMS pcr... Is it dependable to detect SNP using this method?