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Old 12-22-2015, 09:28 AM   #177
Junior Member
Location: Bangkok

Join Date: Sep 2015
Posts: 3

Hi Simone,

I'm trying to amplify one gene of interest; however, I couldn't get a specific band when I added the whole 10 ul of the RT reaction (final PCR volume was 25 ul). The band only appeared when I added 1 ul of the RT mix, suggesting that there's some inhibition of PCR by the RT mix.

However, there was one deviation from your protocol: I used the component version (not ReadyMix) of the KAPA HiFi Hot Start. Do you think that this could have been a factor?

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