Small RNA library contaminated by primers?

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3.5 years ago

eva.lucarelli ▴ 10

Hello,

I have to analyze differential expression on 2 SRA files containing data from a small RNA sequencing experiment (single-ended).

I converted those to .fastq files to control their quality using FastQC, and I get many warnings and errors:

Error in Per base sequence content
Warning in Per sequence GC content
Error in Sequence Duplication Levels
Error in Overrepresented Sequences: sooo many RNA PCR primers indexes (1, 43, 30, 7), and Illumina Small RNA adapter.
Error in Adapter Content: it was expected, Illumina Small RNA 3' adapter.

I used the following line of code to trim the adapter:

trim_galore /root/SRR12737828_pass.fastq.gz /root/SRR12737830_pass.fastq.gz > report

It successfully removed the adaptor, but it looks like it truncated the primers sequence too! I also don't know how to get rid (and if I have to?) of the primers... Am I missing something here?

Thanks in advance for helping me.

Eva

RNA-Seq FastQC primer trimming adapter trimming • 722 views

ADD COMMENT • link 3.5 years ago by eva.lucarelli ▴ 10

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Small RNA data requires special handling. Find out what kit was used for the data. Then follow up with specific instructions for that kit/method.

ADD REPLY • link 3.5 years ago by GenoMax 141k

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