Question: (Closed) RNA-seq from Nanopore featureCounts
0
gravatar for v.shapovalova1
10 months ago by
v.shapovalova110 wrote:

Hello,

I have RNA-seq reads from Nanopore(kit RNA002, U-based fastq), I aligned them with minimap2 and tried to count with featureCounts. The reference was transcriptome from https://www.gencodegenes.org/human/. my command was

 fc<- Rsubread::featureCounts(files="aln.bam",annot.inbuilt="hg38",isLongRead="True")

 Process BAM file aln.bam...                                                ||
||    Single-end reads are included.                                          ||
||    Total alignments : 2081002                                              ||
||    **Successfully assigned alignments : 0 (0.0%)**                             ||
||    Running time : 0.26 minutes   
**Warning message:
In paste("readSummary", ann, files_C, fout, as.numeric(isPairedEnd),  :
  NAs introduced by coercion**

I checked my bam file and have such a statistics:

SN raw total sequences: 632940
SN filtered sequences: 0
SN sequences: 632940
SN is sorted: 1
SN 1st fragments: 632940
SN last fragments: 0
SN reads mapped: 609258
SN reads mapped and paired: 0 # paired-end technology bit set + both mates mapped
SN reads unmapped: 23682

What could it be?

A lot of thanks.

ADD COMMENTlink modified 10 months ago by Devon Ryan96k • written 10 months ago by v.shapovalova110

Please use following option to see if it helps since you have long reads :

  -L                  Count long reads such as Nanopore and PacBio reads. Long
                      read counting can only run in one thread and only reads
                      (not read-pairs) can be counted. There is no limitation on
                      the number of 'M' operations allowed in a CIGAR string in
                      long read counting.
ADD REPLYlink modified 10 months ago • written 10 months ago by genomax89k

Hello v.shapovalova1!

We believe that this post does not fit the main topic of this site.

Cross-posted: https://support.bioconductor.org/p/126571/ Lets keep information focused in that BioC question as the developers are active over there.

For this reason we have closed your question. This allows us to keep the site focused on the topics that the community can help with.

If you disagree please tell us why in a reply below, we'll be happy to talk about it.

Cheers!

ADD REPLYlink written 10 months ago by ATpoint38k
Please log in to add an answer.
The thread is closed. No new answers may be added.

Help
Access

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.
Powered by Biostar version 2.3.0
Traffic: 1607 users visited in the last hour