Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • BBDuk quality filtering not producing expected result

    I'm trying to trim/filter low quality reads from paired-end exome-seq data, using BBDuk.

    I used the command:

    ```
    for ea in $files;
    do
    R1="$ea"
    R2=$(echo $R1 | sed "s/R1/R2/")
    /home/shared/programs/bbmap/bbduk.sh -Xmx1g in1=$R1 in2=$R2 \
    out1="$(echo $ea | sed s/.fastq.gz/_trimmed_filtered.fastq.gz/)" \
    out2="$(echo $(echo $ea | sed s/R1/R2/) | sed s/.fastq.gz/_trimmed_filtered.fastq.gz/)" \
    ref=/home/shared/programs/bbmap/resources/adapters.fa \
    t=10 ktrim=r k=23 kmin=11 hdist=1 maq=10 minlen=60 tpe tbo
    done;
    ```

    After running fastqc on the output of this, I'm seeing that R2 files have some reads with low quality scores (see per sequence quality score), and the overrepresented sequence "NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN".

    Looking at these reads in the fastq:
    ```
    @HISEQ:525:HMFYNBCXX:1:1101:1380:2167 2:N:0:CAGATC
    NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
    +
    !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
    @HISEQ:525:HMFYNBCXX:1:1101:1276:2219 2:N:0:CAGATC
    NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
    +
    !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
    @HISEQ:525:HMFYNBCXX:1:1101:1238:2328 2:N:0:CAGATC
    NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
    ```

    Shouldn't these reads have been filtered out?


    Any help here would be much appreciated.
    Last edited by reliscu; 10-13-2021, 09:15 AM.

Latest Articles

Collapse

  • seqadmin
    Essential Discoveries and Tools in Epitranscriptomics
    by seqadmin


    The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist on Modified Bases...
    Today, 07:01 AM
  • seqadmin
    Current Approaches to Protein Sequencing
    by seqadmin


    Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
    04-04-2024, 04:25 PM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by seqadmin, 04-11-2024, 12:08 PM
0 responses
37 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 10:19 PM
0 responses
41 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 09:21 AM
0 responses
35 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-04-2024, 09:00 AM
0 responses
54 views
0 likes
Last Post seqadmin  
Working...
X