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Tagmentase Adapters

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  • Tagmentase Adapters

    I have a general question about the tagmentase reaction. Has anyone experimented with using single-adapter tagmentase complexes in pairs? For e.g. instead of charging the transposon with an "A" and "B" primer set, making an independent set of Tn5's charged only with primer A and an independent set charged with primer B: then mixing equal amts of the two for the tagmentase reaction? The original shendure paper uses mixed pairs but interestingly the figures provided by illumina show independent complexes as I describe above.

    Also, during the tagmentase reaction, how do you select against two primer A's being adjacent to each other? The enrichment PCR seems like it would amplify both A and B pairs that are adjacent as well as A and A pairs unless the nicking is directional. Thanks in advance for any tips and sorry if this is a novice question, couldn't find specific info within the forums.

  • #2
    to my knowledge, if A and B were added to the same fragment, it will not be amplified or amplified with low efficiency due to the formation of pot shape (suppression PCR).