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Old 11-26-2019, 11:07 AM   #2
Location: Canada

Join Date: Jun 2013
Posts: 54

My guess is that you cannot use the "default" sequencing primers with your scheme.

You appear to have directly attached your gene-specific primer sequence directly to the i5 and i7 illumina indexing primers and therefore lack the sequencing primer binding site, which is actually contained within the Nextera transposase sequence.

Pages 3 and 4 of the following Illumina document might help you:
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