Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • maplesword
    Junior Member
    • May 2010
    • 3

    How to get the exact mismatch positions in Tophat output

    I ran TopHat 1.0.14 and got the mapping results. Now I'm trying to find out which positions are mismatches when mapping to the reference genome for each read. The sam output of TopHat is like this:

    Code:
    [ID]  16  chr1  14509  0  76M  *  0  0  [Query_Seq]  [Quality_Score]  NM:i:2  NH:i:7  CC:Z:chr12  CP:i:91041
    I checked the format manual of SAM. "NM" in TAG part shows the number of differences between the query sequence and the reference sequence. That means in my example there are two mismatches. However, I cannot find out which two positions of this read are the exact mismatch ones in this output. In SAM format, there is one kind of TAG "MD" which shows the mismatch positions but I cannot see any in the output of TopHat. Can anyone tell me how to do?

    Thank you very much!
  • maplesword
    Junior Member
    • May 2010
    • 3

    #2
    I think I have at least solved parts of the problem with samtools.
    First I used "import" command of samtools to transform the sam file to bam format, and then used "fillmd" command to add the MD tag.

    But it comes another problem. When adding the MD tag, the program also output lots of warnings showing that the NM tags of lots of reads are wrong. I don't know why. Can anyone give me help? Thank you!

    Comment

    Latest Articles

    Collapse

    • SEQadmin2
      Advanced Sequencing Platforms Tackle Neuroscience’s Toughest Genomics Problems
      by SEQadmin2



      Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
      ...
      07-09-2026, 11:10 AM
    • SEQadmin2
      Cancer Drug Resistance: The Lingering Barrier to Rising Survival
      by SEQadmin2



      Cancer survival rates have significantly increased in the last few decades in the United States, reaching a combined 70% 5-year survival rate by 2021. Behind this number, there are years of research to find new therapies, drug targets, and early detection methods. But there is one core challenge that keeps slowing down these advances, and it’s about drug resistance.

      There is no single reason why many patients don’t respond to treatment as expected. Cancer is...
      07-08-2026, 05:17 AM
    • GATTACAT
      Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
      by GATTACAT
      Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
      07-01-2026, 11:43 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by SEQadmin2, Today, 10:26 AM
    0 responses
    9 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-09-2026, 10:04 AM
    0 responses
    24 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-08-2026, 10:08 AM
    0 responses
    16 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-07-2026, 11:05 AM
    0 responses
    33 views
    0 reactions
    Last Post SEQadmin2  
    Working...