Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • bioramg
    Member
    • Apr 2014
    • 12

    SPAdes assembler

    Hello,
    I tried to assemble a mitochondrial genome assembly with SPAdes assembler using pair-end Illumina and Nanopore sequencing data. The size of the Illumina data is 18 GB and when I do hybrid assembly after trimming adapters, it showing that Memory is insufficient to run this program and requires at least 389 GB RAM. But, I am having 188 GB RAM.
    Then, I used BBmerge commands to sort out this issue. But when I run pair-end reads using bbmerge-auto.sh command, its saying that "try increasing the -Xmx flag and using tool specific memory related parameters".

    Please suggest me how to do hybrid assembly with both Illumina and Nanopore data?
  • SNPsaurus
    Registered Vendor
    • May 2013
    • 525

    #2
    Is your 18 Gb of Illumina data pure mtDNA or mtDNA plus nuclear? If it is pure mtDNA then you probably have many thousand-fold coverage of the mitochondrial genome and you can reduce the number of reads used as input (bbtools reformat will do that if you give the paired-end reads as in= and in2= and then use reads=10000000 or some other number). If it is not pure mtDNA then you probably still have high coverage and could cut the input by 3/4s.
    Providing nextRAD genotyping and PacBio sequencing services. http://snpsaurus.com

    Comment

    • bioramg
      Member
      • Apr 2014
      • 12

      #3
      Thank you so much for your kind reply.

      Its whole genomic data that includes, chloroplast, mitochondrial and nuclear genome data. How can I choose 3/4 read? Could you please suggest.

      Thank you so much.

      With warm regards,
      Raman. G

      Comment

      • SNPsaurus
        Registered Vendor
        • May 2013
        • 525

        #4
        I would use bbtools reformat to set the number of desired reads.
        Providing nextRAD genotyping and PacBio sequencing services. http://snpsaurus.com

        Comment

        • bioramg
          Member
          • Apr 2014
          • 12

          #5
          Could you please give me with some examples. I am new to bioinformatics. How to set desired reads...

          Comment

          • SNPsaurus
            Registered Vendor
            • May 2013
            • 525

            #6
            You used bbmerge already, so it sounds like you have bbtools installed.
            reformat.sh in=read1,fq,gz in2=read2.fq.gz out=read1_fewer.fq.gz out2=read2_fewer.fq.gz reads=10000000
            Providing nextRAD genotyping and PacBio sequencing services. http://snpsaurus.com

            Comment

            • bioramg
              Member
              • Apr 2014
              • 12

              #7
              Thank you so much.

              by the way, I have used this command to estimate genome size using the following command:
              java -ea -Xmx188g -Xms188g -cp /home/pmslab/Desktop/Raman/bin/bin/bbmap/current/ jgi.KmerCountExact in=trim_CKEMJ1.fastq in2=trim_CKEMJ2.fastq khist=khist_trim_CKEMJ.txt peaks=peaks_trim_CKEMJ.txt -Xmx188g

              But i got an error message:
              ./../bin/bin/bbmap/kmercountexact.sh: line 138: 30199 Killed

              What could be the problem?
              Thank you.

              Comment

              Latest Articles

              Collapse

              • SEQadmin2
                Nine Things a Sample Prep Scientist Thinks About Before Sequencing
                by SEQadmin2


                I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

                Here are nine questions we think about, in roughly the order they matter, before...
                06-18-2026, 07:11 AM
              • SEQadmin2
                From Collection to Sequencing: Why Sample Preparation and Preservation Define Sequencing Data
                by SEQadmin2


                Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.


                The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
                ...
                06-02-2026, 10:05 AM

              ad_right_rmr

              Collapse

              News

              Collapse

              Topics Statistics Last Post
              Started by SEQadmin2, 06-26-2026, 11:10 AM
              0 responses
              10 views
              0 reactions
              Last Post SEQadmin2  
              Started by SEQadmin2, 06-17-2026, 06:09 AM
              0 responses
              45 views
              0 reactions
              Last Post SEQadmin2  
              Started by SEQadmin2, 06-09-2026, 11:58 AM
              0 responses
              105 views
              0 reactions
              Last Post SEQadmin2  
              Started by SEQadmin2, 06-05-2026, 10:09 AM
              0 responses
              125 views
              0 reactions
              Last Post SEQadmin2  
              Working...