Hi,
Maybe someone has faced the same problem, as I am:
I am using Agencourt AMPure XP to clean my PCR-amplicons from PCR-reaction mixture. I am using AMPure at 1x sample volume, to get rid of all the primers.
In the last few months I started noticing, that the AMPure magnetic beads do not pellet properly on the tube wall after the addition of Elution Buffer (10 mM Tris-HCl, pH 8,5). The DNA concentrations of amplicons also decreased. I tried to elute my amplicons in MQ water and it worked well, I also obtained a bit higher DNA concentrations. But I am still wondering, what is wrong with the elution using EB. I checked my Ethanol and tried to use 80% instead of 70%, it did not change anything. I also tried to use different lab plastic, magnet plate and ordered fresh EB and AMPure. Nothing worked and I still got this „haze“ instead of properly pelleted beads.
Maybe someone could help me, what am I doing wrong?
Maybe someone has faced the same problem, as I am:
I am using Agencourt AMPure XP to clean my PCR-amplicons from PCR-reaction mixture. I am using AMPure at 1x sample volume, to get rid of all the primers.
In the last few months I started noticing, that the AMPure magnetic beads do not pellet properly on the tube wall after the addition of Elution Buffer (10 mM Tris-HCl, pH 8,5). The DNA concentrations of amplicons also decreased. I tried to elute my amplicons in MQ water and it worked well, I also obtained a bit higher DNA concentrations. But I am still wondering, what is wrong with the elution using EB. I checked my Ethanol and tried to use 80% instead of 70%, it did not change anything. I also tried to use different lab plastic, magnet plate and ordered fresh EB and AMPure. Nothing worked and I still got this „haze“ instead of properly pelleted beads.
Maybe someone could help me, what am I doing wrong?
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