Hi all!
Did anyone try that method yet? I’m a little confused by the paper, the main advantage of that variable length primer seems to be a reduction of PhiX spike in. However in the Kozich et al. paper you guys use less than 10% PhiX anyway, and it seems to work fine. Fadrosh et al. say the general quality of the sequences improves with their method also, apart from the PhiX issue. Do you think this is likely? There does not seem to be a direct comparison in the paper.
Cheers,
Till
Fadrosh, D.W., Ma, B., Gajer, P., Sengamalay, N., Ott, S., Brotman, R.M., and Ravel, J. (2014). An improved dual-indexing approach for multiplexed 16S rRNA gene sequencing on the Illumina MiSeq platform. Microbiome 2, 1–7.