I am another newbie trying to get through a first project in NGS.
We are trying to amplify universal prokaryotic sequences for 16s rRNA gene (V3-V4 region) from benthic marine critters - mostly from microbiome extractions using materials from invertebrates and some algae. I was told that using the Fadrosh et al. 2014 dual-indexing method would improve our results a lot when we do PE250. Hopefully that is true - yes? (No?)
Also, right now all the current primer sets are going through continued evaluation and most seem to be found lacking in one way or another. Does anyone have a strong opinion on choices based on actual experience with getting good results? Has anyone tried the primers discussed in Takahashi et al. 2014, Pro341F/Pro805R? Did they work as well as claimed?
Thanks for any help pointing me in the right direction!
We are trying to amplify universal prokaryotic sequences for 16s rRNA gene (V3-V4 region) from benthic marine critters - mostly from microbiome extractions using materials from invertebrates and some algae. I was told that using the Fadrosh et al. 2014 dual-indexing method would improve our results a lot when we do PE250. Hopefully that is true - yes? (No?)
Also, right now all the current primer sets are going through continued evaluation and most seem to be found lacking in one way or another. Does anyone have a strong opinion on choices based on actual experience with getting good results? Has anyone tried the primers discussed in Takahashi et al. 2014, Pro341F/Pro805R? Did they work as well as claimed?
Thanks for any help pointing me in the right direction!
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