Help with PICRUSt Analysis Using Nanopore Metabarcoding Data

[Ana Benavides]

Hello everyone,

I am working with metabarcoding data obtained through Nanopore sequencing and need assistance with conducting functional prediction analysis using PICRUSt. Here are the details of my situation and what I have done so far:

Project Details:

• I have sequenced 16S rRNA samples using a Nanopore platform.
• I have completed taxonomic assignment using Kraken2.

Steps I have taken:

1. I have the FASTQ files and the taxonomic assignment results from Kraken2.

I need help with:

1. How to convert and prepare my FASTQ files to be compatible with QIIME 2 to obtain ASVs.
2. How to export the necessary files from QIIME 2 in a format compatible with PICRUSt.
3. The exact steps to run PICRUSt with my data, considering it originates from a Nanopore platform.
4. Any advice on specific adjustments I should make to ensure the accuracy of the results.