We've been struggling to get fusion primers to work on IT system for 16s tag sequencing. Primary reason for using the system--don't do much genomics. Don't know why it took so long to check this, but it looks like the problem is not the fusion primers but the IT sequences themselves (see attached). So these work fine in adapter ligation, but not in PCR. We get faint, broad double bands, generally one at expected size and other higher. Not enough material to size select or even simply clean up.
Anyone else trying this approach? Clearly the 5' ends are going to have to be kept for binding to spheres, but maybe we can change the sequencing primer complementary part--?
Ion Torrent folks, please chime in...will also post to Ion Community.
Anyone else trying this approach? Clearly the 5' ends are going to have to be kept for binding to spheres, but maybe we can change the sequencing primer complementary part--?
Ion Torrent folks, please chime in...will also post to Ion Community.
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