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.
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.
Comment