#16 Abundance file bug

[Anonymous]

Hello,

I am trying to create a Illumina 16S amplicon library from V3V4 variable regions with certain abundance for each reference. This library is an overlaping paired-end one. I used the following command:

grinder -reference_file 16s_seq_example -total_reads 1000 -read_dist 300 -insert_dist 450 -mo FR -forward_reverse 16Sprimers.fna -md poly4 3e-3 3.3e-8 -abundance_file abun_example -qual_levels 30 30 -fastq_output 1

This is the message that the program retrieved me:

Error: Requested reference sequence 'AE000666' in file 'abundance_file.txt' does not exist in the input database.

I attach the files that I am using as input.

[Anonymous]

Sorry, I didn's save the abundance and reference files in my last message.

The abundance file contains the following lines:

AE000666 50.0
AE003849 50.0

[floflooo]

Hi,
The sofware is correct. It does not find reference sequence AE000666 because it is not amplified by the primers you provided. You need to not request this sequence or modified your primers so that they amplify it.
Best,
Florent

[Anonymous]