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Old 08-10-2017, 07:16 PM   #1
Buckethead84
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Location: New York

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Default AmpliSeq Design

Could someone explain to me how AmpliSeq for Ion Torrent works? I understand how it's a multiplex PCR, but I've heard that it uses uracil containing primers that they aid in the sensitivity and specificity of the PCR reactions because the RNA/DNA interactions are much stronger than DNA/DNA interactions. This is potentially why the ends of the amplicons need to be digested for the rest of the library preparation to work? Is there any truth to this?

If that's true, could I use something like a polymerase that isn't inhibited by uracil to convert AmpliSeq amplicons into Illumina compatible amplicons?

I would really appreciate and resources anyone has... I feel like this stuff is exceptionally hard to find.
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