Hello @Rainjie,
I utilized 'barrnap' to extract the 16S rRNA sequences from the MAGs I assembled in the metagenome analysis. Surprisingly, I noticed the presence of multiple 16S rRNA sequences within the same MAGs, which appears to contradict my understanding of 16S amplification data. In this situation, how should I determine the appropriate 16S rRNA sequence for each MAG?"
These multiple 16S sequences, are they taxonomically distinct, or are they just copies of one another? Some microbes have more than one copy of the 16S gene in their genome.
Guidance in needed on incorporating the 16S sequences represented by the obtained MAGs into the qiime2 database. Specifically, I intend to add these sequences to the Sliva 138 database. Could you please advise me on the appropriate steps to achieve this or should I try
RESCRIPt?
What do you hope to accomplish by incorporating the mag-extracted 16S sequences into a classification database? It would possible to add your own sequences to the Silva 138 database sequences and re-train, but I'm not sure how useful that would be.