[Hannah Seidel]

I am interested in using this protocol with AMPure XP beads. My goal is to clean-up PCR reactions for Sanger sequencing at less cost and less hands-on time than clean-ups using silica columns.

The AMPure literature says that I should add ~2 volumes their beads to 1 volume of PCR reaction. Their literature also says that 5 uL beads is more than enough to bind any reasonable quantity of DNA.

So, my plan is clean-up my PCR reactions by adding 5 uL AMPure XP beads and ~2 volumes BOMB binding buffer (prepared without beads) to 1 volume PCR reaction (50 uL, typically). This protocol will allow me to reduce the amount of AMPure XP beads by 20-fold and thus save money.

Do you think my idea will work?

Do I need to “wash” the AMPure XP beads first? BOMB protocol 4.2 says that carboxyl-coated beads should be washed before preparing the binding buffer. What do you wash them in? The protocol gives no instructions for the wash step.

Thanks for your help!

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