Dear Qiime2 users, I'm trying to use paired-end sequences downloaded from SRA, but I have problems with the command, for example:
**SRA files (one per sample): i.e., experiment SRX2730405 (view SRX2730405.txt (3.3 KB)
**Manifest file: Map-imput_Narish2017.csv (164 Bytes)
** comand
qiime tools import
--type 'SampleData[PairedEndSequencesWithQuality]'
--input-path Map-imput_Naris2017.csv
--output-path Narish2017-demux.qza
--input-format PairedEndFastqManifestPhred64
I already tried several forms of --type, and --input-format, but did not identify the problem. Please, help me identify and correct the error.
Also, I would appreciate any recommendation on how to download data from SRA (I just identified and downloaded each experiment as .fastq
I suspect the issue is in the manifest file. I don’t see a file extension listed as part of the path. I would expect those paths to end in .fastq (or maybe .fastq.gz). If you run the following command, what do you see?
ls /mnt/c/Dowload_linux/set-sequence/
Otherwise, could you post what error you are seeing specifically?
I see that the sequences R1 and R2 are in the same file (SRA run), is it necessary to separate them in order to import them?
I was reading about FASTQ de-interlacer on paired-end reads (on galaxy)...what do you suggest?
cheers
Hey there @Dasiel! It looks like there might be an issue related to the manifest file itself. I noticed you are running an older version of QIIME 2 (2018.8). If you upgrade to 2018.11 you will see a more detailed error message about the manifest file that can hopefully set you in the right direction.
One thing I notice is that the filename in your manifest says "SRX2730405_1.fastq", but the closest filename in the dir is called "SRX2730405.fastq" (note the missing _1 at the end).
Hello Matthew, thanks for your observation. Normally I work on the lab server, and we have the current version, but I will also update my laptop, thanks.
The sequences are interlaced, what do you recommend me?
Unfortunately we don’t have a mechanism in QIIME 2 for dealing with interlaced reads — if you are able to deinterlace using an external tool (see this link for a suggestion) you could then import those deinterlaced reads using the manifest format you started working with above. Sorry!
Great, in the exchange with you I can understand the problem!. Interesting that not all the samples (run) in this set are interlaced, so I’m checking them individually. I’m using FASTQ de-interlacer (Galaxy version). Thanks for your important support to the users of qiime2. Cheers
