Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Variants called using GATK from samples of Nextera and TruSeq exome enrichment kits

    I am doing variant calling for exomes generated from Illumina HiSeq 2000, with Nextera Exome Enrichment kit.
    Earlier I used the same variant calling pipeline to call variants from sequences based on TruSeq Exome Enrichment kit.
    Now with the Nextera Exome Enrichment kit, I find that the number of variants called using GATK is reduced to almost half as compared to TruSeq.
    I am not able to reach a conclusion as to why this drastic reduction in number.

    TruSeq sample Nextera sample
    Mean coverage 40X 47.5X
    GATK variants 53917 27596

    Here is the GATK command I used for calling variants for both TruSeq and Nextera samples:
    java -Xmx2g -Djava.io.tmpdir=temp -jar $gatk_jar -T UnifiedGenotyper -I $in -R $ucsc_hg19_fasta -D:name,VCF $dbsnp146_hg19_vcf -o $o -dcov 1000 -A AlleleBalance -A BaseCounts -A VariantType -baq CALCULATE_AS_NECESSARY -stand_call_conf 30.0 -stand_emit_conf 10.0 -glm BOTH -L $nextera_target_bed(or $truseq_target_bed )

  • #2
    A few possible explanations:

    1) higher error rate in the sequence data from the TruSeq sample, producing false-positive variant calls. You can use FastQC to assess the base quality metrics of the two data sets.

    2) capture of highly polymorphic loci in the TruSeq but not Nextera enrichment. Compare the covered intervals with Bedtools, and see if there's a large difference in variant frequency between the shared vs unique regions.

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Recent Advances in Sequencing Analysis Tools
      by seqadmin


      The sequencing world is rapidly changing due to declining costs, enhanced accuracies, and the advent of newer, cutting-edge instruments. Equally important to these developments are improvements in sequencing analysis, a process that converts vast amounts of raw data into a comprehensible and meaningful form. This complex task requires expertise and the right analysis tools. In this article, we highlight the progress and innovation in sequencing analysis by reviewing several of the...
      Yesterday, 07:48 AM
    • 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...
      04-22-2024, 07:01 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, Yesterday, 07:17 AM
    0 responses
    11 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 05-02-2024, 08:06 AM
    0 responses
    19 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-30-2024, 12:17 PM
    0 responses
    20 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-29-2024, 10:49 AM
    0 responses
    28 views
    0 likes
    Last Post seqadmin  
    Working...
    X