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Porechop demulitplexing of nanopore sequencing data and barcode bins for each sample

Dear, I am new to nanopore community and have some questions. I might sound silly! We preapred nanopore librariy using 1-24 SQK-16S024 kit for 24 samples. After sequencing, i got fast5 and fastq files. Fastq files were subjected to Porechop for demultiplexing and adapter trimming. As an output, porechop created only 2 barcode bins, BC01 and BC02. BC01 is a very big file with more than 1 million reads. The question is how many barcode bins were expected after porechop process as we created 24 sample library? I was expecting 24 barcode bins as i used 24 samples for the library preparation. COuld you please share your experience, it might be helpful. Thanks!

porechop
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updated by samuel.a.odonnell samuel.a.odonnell • written by muhammad.akram • muhammad.akram

While the kit is for 24 samples, how many barcodes did you actually use?

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written by andres.firrincieli andres.firrincieli

OP says:

I was expecting 24 barcode bins as i used 24 samples for the library preparation.

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written by GenoMax GenoMax

Considering you barcoded 24 samples, you should expect 24 barcode bins as you stated.

I am not sure porechop has the barcodes for the kit you have used. This might be your issue. You should try using the demultiplexing in guppy instead anyway

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written by samuel.a.odonnell samuel.a.odonnell

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